Dual pharmacophores—PDE4-muscarinic antagonistics

ABSTRACT

The present invention is directed to novel compounds of Formula (I), pharmaceutical compositions and their use in therapy, for example as inhibitors of phosphodiesterase type IV (PDE4) and as antagonists of muscarinic acetylcholine receptors (mAChRs), in the treatment of/and or prophylaxis of respiratory diseases, including antiinflammatory and/or allergic diseases such as chronic obstructive pulmonary disease (COPD), asthma, rhinitis (e.g. allergic rhinitis), atopic dermatitis or psoriasis.

CROSS REFERENCE TO PRIOR APPLICATIONS

This application claims the benefit of U.S. Provisional application 61/026,572 filed 6 Feb. 2008.

FIELD OF THE INVENTION

The present invention relates to novel compounds of Formula (I), or salts thereof, processes for their preparation, intermediates usable in these processes, and pharmaceutical compositions containing the compounds or salts. The invention also relates to the use of these compounds or salts thereof in therapy, for example as inhibitors of phosphodiesterase type IV (PDE4) and as antagonists of muscarinic acetylcholine receptors (mAChRs), and useful in the treatment of, and/or prophylaxis of respiratory diseases, including anti-inflammatory and allergic diseases such as chronic obstructive pulmonary disease (COPD), asthma, rhinitis (e.g. allergic rhinitis), atopic dermatitis or psoriasis.

BACKGROUND OF THE INVENTION

Acetylcholine released from cholinergic neurons in the peripheral and central nervous systems affects many different biological processes through interaction with two major classes of acetylcholine receptors—the nicotinic and the muscarinic acetylcholine receptors. Muscarinic acetylcholine receptors (mAChRs) belong to the superfamily of G-protein coupled receptors having seven transmembrane domains. There are five subtypes of mAChRs, termed M1-M5, and each is the product of a distinct gene. Each of these five subtypes displays unique pharmacological properties. Muscarinic acetylcholine receptors are widely distributed in vertebrate organs, and these receptors can mediate both inhibitory and excitatory actions. For example, in smooth muscle found in the airways, M3 mAChRs mediate contractile responses. For a review, see Caulfield (1993 Pharmac. Ther. 58:319-79).

In the lungs, mAChRs have been localized to smooth muscle in the trachea and bronchi, the submucosal glands, and the parasympathetic ganglia. Muscarinic receptor density is greatest in parasympathetic ganglia and then decreases in density from the submucosal glands to tracheal and then bronchial smooth muscle. Muscarinic receptors are nearly absent from the alveoli. For review of mAChR expression and function in the lungs, please see Fryer and Jacoby (1998 Am J Respir Crit Care Med 158(5, pt 3) S 154-60).

Three subtypes of mAChRs have been identified as important in the lungs, M1, M2 and M3 mAChRs. The M3 mAChRs, located on airway smooth muscle, mediate muscle contraction. Stimulation of M3 mAChRs activates the enzyme phospholipase C via binding of the stimulatory G protein Gq/11 (Gs), leading to liberation of phosphatidyl inositol-4,5-bisphosphate, resulting in phosphorylation of contractile proteins. M3 mAChRs are also found on pulmonary submucosal glands. Stimulation of this population of M3 mAChRs results in mucus secretion.

M2 mAChRs make up approximately 50-80% of the cholinergic receptor population on airway smooth muscles. Although the precise function is still unknown, they inhibit catecholaminergic relaxation of airway smooth muscle via inhibition of cAMP generation. Neuronal M2 mAChRs are located on postganglionic parasympathetic nerves. Under normal physiologic conditions, neuronal M2 mAChRs provide tight control of acetylcholine release from parasympathetic nerves. Inhibitory M2 mAChRs have also been demonstrated on sympathetic nerves in the lungs of some species. These receptors inhibit release of noradrenaline, thus decreasing sympathetic input to the lungs.

M1 mAChRs are found in the pulmonary parasympathetic ganglia where they function to enhance neurotransmission. These receptors have also been localized to the peripheral lung parenchyma, however their function in the parenchyma is unknown.

Muscarinic acetylcholine receptor dysfunction in the lungs has been noted in a variety of different pathophysiological states. In particular, in asthma and chronic obstructive pulmonary disease (COPD), inflammatory conditions lead to loss of inhibitory M2 muscarinic acetylcholine autoreceptor function on parasympathetic nerves supplying the pulmonary smooth muscle, causing increased acetylcholine release following vagal nerve stimulation (Fryer et al. 1999 Life Sci 64 (6-7) 449-55). This mAChR dysfunction results in airway hyperreactivity and hyperresponsiveness mediated by increased stimulation of M3 mAChRs.

Recent literature has focused on the non-neuronal cholinergic system in the lungs where there is an emerging literature supporting a role for muscarinic receptors in mediating immunomodulatory and inflammatory functions in respiratory diseases such as asthma and COPD. Many of the components for cholinergic signaling have been reported to be contained within inflammatory and resident cells of the lungs, including muscarinic receptor expression on lymphocytes, alveolar macrophages, mast cells and epithelial cells. The view that acetylcholine is solely a neurotransmitter of the parasympathetic nervous system is currently being challenged as there is mounting evidence to suggest it has an integral role in host defense and airway inflammation. For a full review see Gwilt et al., 2007 (Gwilt CR. et al., The non-neuronal cholinergic system in the airways: An unappreciated regulatory role in pulmonary inflammation? Pharmacol. Ther. 2007; 115: 208-222) and Kummer & Lips 2006 (Kummer W and Lips KS. Non-neuronal acetylcholine release and its contribution to COPD pathology. Drug Discovery Today: Disease Mechanisms 2006; 3:47-52). A consequence of this emerging science is the implication that anti-cholinergic antagonists may have a much broader therapeutic potential for respiratory diseases with anti-inflammatory and disease modifying activity as well as the their well established utility as bronchodilator agents.

COPD is an imprecise term that encompasses a variety of progressive health problems including chronic bronchitis, chronic bronchiolitis and emphysema, and it is a major cause of mortality and morbidity in the world. Smoking is the major risk factor for the development of COPD; nearly 50 million people in the U.S. alone smoke cigarettes, and an estimated 3,000 people take up the habit daily. As a result, COPD is expected to rank among the top five as a world-wide health burden by the year 2020. Inhaled anti-cholinergic therapy is currently considered the “gold standard” as first line therapy for COPD (Pauwels et al. 2001 Am. J. Respir. Crit. Care Med. 163:1256-1276).

Despite the large body of evidence supporting the use of anti-cholinergic therapy for the treatment of airway hyperreactive diseases, relatively few anti-cholinergic compounds are available for use in the clinic for pulmonary indications. Ipratropium Bromide (Atrovent®; and Combivent®, in combination with albuterol) is one of the few inhaled anti-cholinergic marketed for the treatment of airway hyperreactive diseases. While this compound is a potent anti-muscarinic agent, it is short acting, and thus must be administered as many as four times daily in order to provide relief for the COPD patient. The long-acting anti-cholinergic Tiotropium Bromide (Spiriva®) has recently been approved in a number of countries.

Since mAChRs are widely distributed throughout the body, the ability to apply anti-cholinergics locally and/or topically to the respiratory tract is particularly advantageous, as it would allow for lower doses of the drug to be utilized. Furthermore, the ability to design topically active drugs that have long duration of action, and in particular, are retained either at the receptor or by the lung, would allow the avoidance of unwanted side effects that may be seen with systemic anti-cholinergic use.

WO 2004/091482 describes a dimeric bicyclic amine derivative having anti-muscarinic receptor activity:

wherein, inter alia, X represents a group of the formula (d) or (e): —Y—Ar—Y—  (d) —Y-L-Y—  (e)

Y is selected from the group consisting of a bond, OR², SR², NR²R³, and C₁₋₄ alkyl; and L represents a bond, C₁₋₄ alkyl or C₃₋₈ cycloalkyl.

WO 2005/095407 also discloses a similar dimeric bicyclic amine derivative to that above having anti-muscarinic receptor activity wherein inter alia, X is a group of the formula (d), (e) and (f): —Y—Ar—Y—  (d) —Y-L-Y—  (e) Y—Ar¹—Z—Ar²—Y  (f)

Y is, independently, selected from the group consisting of a bond, O, S, NR², —NR²C₁₋₄ alkyl-, and C₁₋₄ alkyl-; each of the alkyl groups may contain a heteroatom selected from O, NR², or S; and

Z represents a bond, O, NR², S, C₁₋₄ alkylidene or C₁₋₄ alkyl.

Other mAChR antagonists, non-dimeric in structure, may be found in WO 2004/012684; WO 2005/009439; WO 2005/09362; WO 2005/09440; WO 2005/037280; WO 2005/037224; WO 2005/046586; WO 2005/055940; WO 2005/055941; WO 2005/067537; WO 2005/087236; WO 2005/086873; WO 2005/094835; WO 2005/094834; WO 2005/094251; WO 2005/099706; WO 2005/104745; WO 2005/112644; WO 2005/118594; WO 2006/005057; WO 2006/017767; WO 2006/017768; WO 2006/050239; WO 2006/055503; WO 2006/055553; WO 2006/062931; WO 2006/062883; WO 2006/065788; WO 2006/065755; WO 2007/018514; WO 2007/018508; WO 2007/016639; WO 2007/016650; and WO 2007/022351.

NVA237 (glycopyrrolate) glycopyrrolate or glycopyrronium bromide, a quaternary ammonium derivative with anticholinergic and antimuscarinic properties. It is being developed by Novartis for once daily treatment of COPD.

LAS-34273, also known as aclidinium bromide, is a quaternary ammonium anticholinergic muscarinic M3 antagonist originated by Almirall and believed to be in phase 3 development for treating COPD.

With respect to the PDE4 moieties: U.S. Pat. Nos. 3,979,399, 3,840,546, and 3,966,746 (E.R.Squibb & Sons) disclose 4-amino derivatives of pyrazolo[3,4-b]pyridine-5-carboxamides wherein the 4-amino group NR₃R₄ can be an acyclic amino group wherein R₃ and R₄ may each be hydrogen, lower alkyl (e.g. butyl), phenyl, etc.; NR₃R₄ can alternatively be a 3-6-membered heterocyclic group such as pyrrolidino, piperidino and piperazino. The compounds are disclosed as central nervous system depressants useful as ataractic, analgesic and hypotensive agents.

U.S. Pat. Nos. 3,925,388, 3,856,799, 3,833,594 and 3,755,340 (E.R.Squibb & Sons) disclose 4-amino derivatives of pyrazolo[3,4-b]pyridine-5-carboxylic acids and esters. The compounds are mentioned as being central nervous system depressants useful as ataractic agents or tranquilizers, as having anti-inflammatory and analgesic properties. The compounds are mentioned as increasing the intracellular concentration of adenosine-3′,5′-cyclic monophosphate and for alleviating the symptoms of asthma.

H. Hoehn et al., J. Heterocycl Chem., 1972, 9(2), 235-253 discloses a series of 1H-pyrazolo[3,4-b]pyridine-5-carboxylic acid derivatives with 4-hydroxy, 4-chloro, 4-alkoxy, 4-hydrazino, and 4-amino substituents. Ethyl 4-(n-butylamino)-1-ethyl-1H-pyrazolo[3,4-b]-pyridine-5-carboxylate is disclosed therein; this compound is cartazolate.

The compound tracazolate, ethyl 4-(n-butylamino)-1-ethyl-6-methyl-1H-pyrazolo[3,4-b]-pyridine-5-carboxylate, is known as an anxiolytic agent (e.g. see J.B. Patel et al., Eur. J. Pharmacol., 1982, 78, 323). Other 1-substituted 4-(NH₂ or NH-alkyl)-1H-pyrazolo[3,4-b]-pyridine-5-carboxylic acid esters and amides are disclosed as potential anxiolytic agents in T.M. Bare et al., J. Med. Chem., 1989, 32, 2561-2573.

CA 1003419, CH 553 799 and T. Denzel, Archiv der Pharmazie, 1974, 307(3), 177-186 disclose 4,5-disubstituted 1H-pyrazolo[3,4-b]pyridines unsubstituted at the 1-position.

Japanese laid-open patent application JP-2002-20386-A (Ono Yakuhin Kogyo KK) published on 23 Jan. 2002 discloses pyrazolopyridine compounds of the following inter alia formula:

The compounds of JP-2002-20386-A are stated as having PDE4 inhibitory activity and as being useful in the prevention and/or treatment of inflammatory diseases and many other diseases.

1,3-Dimethyl-4-(arylamino)-pyrazolo[3,4-b]pyridines with a 5-C(O)NH₂ substituent similar or identical to those in JP-2002-20386-A were disclosed as orally active PDE4 inhibitors by authors from Ono Pharmaceutical Co. in: H. Ochiai et al., Bioorg. Med. Chem. Lett., 2004, vol. 14(1), pp. 29-32. Full papers on these and similar compounds as orally active PDE4 inhibitors are: H. Ochiai et al., Bioorg. Med. Chem., 2004, 12(15), 4089-4100, and H. Ochiai et al., Chem. Pharm. Bull., 2004, 52(9), 1098-1104.

EP 0 076 035 A1 (ICI Americas) discloses pyrazolo[3,4-b]pyridine derivatives as central nervous system depressants useful as tranquilizers or ataractic agents for the relief of anxiety and tension states.

J.W. Daly et al., Med. Chem. Res., 1994, 4, 293-306 and D. Shi et al., Drug Development Research, 1997, 42, 41-56 disclose a series of 4-(amino)substituted 1H-pyrazolo[3,4-b]pyridine-5-carboxylic acid derivatives, including ethyl 4-cyclopentylamino-1-methyl-1H-pyrazolo[3,4-b]pyridine-5-carboxylate, and their affinities and antagonist activities at A₁- and A_(2A)-adenosine receptors, and the latter paper discloses their affinities at various binding sites of the GABA_(A)-receptor channel. S. Schenone et al., Bioorg. Med. Chem. Lett., 2001, 11, 2529-2531, and F. Bondavalli et al., J. Med. Chem., 2002, 45(22), pp. 4875-4887 disclose a series of 4-amino-1-(2-chloro-2-phenylethyl)-1H-pyrazolo[3,4-b]pyridine-5-carboxylic acid ethyl esters as A₁-adenosine receptor ligands.

WO 02/060900 A2 appears to disclose, as MCP-1 antagonists for treatment of allergic, inflammatory or autoimmune disorders or diseases, a series of bicyclic heterocyclic compounds with a —C(O)—NR⁴—C(O)—NR⁵R⁶ substituent, including isoxazolo[5,4-b]pyridines and 1H-pyrazolo[3,4-b]pyridines (named as pyrazolo[5,4-b]pyridines) with the —C(O)—NR⁴—C(O)—NR⁵R⁶ group as the 5-substituent and optionally substituted at the 1-, 3-, 4-, and/or 6-positions. Bicyclic heterocyclic compounds with a —C(O)NH₂ substituent instead of the —C(O)—NR⁴—C(O)—NR⁵R⁶ substituent are alleged to be disclosed in WO 02/060900 as intermediates in the synthesis of the —C(O)—NR⁴—C(O)—NR⁵R⁶ substituted compounds. See also WO 02/081463 A1 for similar MCP-1 antagonists.

WO 00/15222 (Bristol-Myers Squibb) discloses inter alia pyrazolo[3,4-b]pyridines having inter alia a C(O)—X₁ group at the 5-position and a group E₁ at the 4-position of the ring system. Amongst other things, X₁ can for example be —OR₉, —N(R₉)(R₁₀) or —N(R₅)(-A₂-R₂), and E₁ can for example be —NH-A₁-cycloalkyl, —NH-A₁-substituted cycloalkyl, or —NH-A₁-heterocyclo; wherein A₁ is an alkylene or substituted alkylene bridge of 1 to 10 carbons and A₂ can for example be a direct bond or an alkylene or substituted alkylene bridge of 1 to 10 carbons. The compounds are disclosed as being useful as inhibitors of cGMP phosphodiesterase, especially PDE type V, and in the treatment of various cGMP-associated conditions such as erectile dysfunction.

H. de Mello, A. Echevarria, et al., J. Med. Chem., 2004, 47(22), 5427-5432, discloses 3-methyl or 3-phenyl 4-anilino-1H-pyrazolo[3,4-b]pyridine 5-carboxylic esters as potential anti-Leishmania drugs.

WO 2004/056823 A1 (PCT/EP2003/014867, filed on 19 Dec. 2003, published on 8 Jul. 2004, Glaxo Group Limited), and incorporated herein by reference in its entirety as though fully set forth, discloses and claims pyrazolo[3,4-b]pyridine compounds or salts thereof with a 4-NR³R^(3a) group (R^(3a) is preferably H) and with a group Het at the 5-position of the pyrazolo[3,4-b]pyridine, wherein Het is usually a 5-membered optionally substituted heteroaryl group.

WO 2004/056823 A1 also discloses the use of these compounds as PDE4 inhibitors and for the treatment and/or prophylaxis of inter alia COPD, asthma or allergic rhinitis.

WO 2004/024728 A2 (PCT/EP2003/011814, filed on 12 Sep. 2003, published on 25 Mar. 2004, Glaxo Group Limited), discloses pyrazolo[3,4-b]pyridine having the following generic formula.

In WO 2004/024728 A2, pyrazolo[3,4-b]pyridine compounds are disclosed as being inhibitors of PDE4. WO 2004/024728 and WO 2004/056823 are noted in Expert Opin. Ther. Patents, 2005 (January edition), 15(1), 111-114.

WO 2005/058892 A1 (PCT/EP2004/014490, filed on 17 Dec. 2004, published on 30 Jun. 2005, Glaxo Group Limited), discloses pyrazolo[3,4-b]pyridine compounds for use as PDE4 inhibitors for treating inflammatory or allergic diseases such as COPD, asthma, rheumatoid arthritis, allergic rhinitis or atopic dermatitis.

Further pyrazolo[3,4-b]pyridine compounds and their use as PDE4 inhibitors, are disclosed in patent applications WO 2005/090353 A1 (PCT/GB2005/000976), WO 2005/090348 A1 (PCT/GB2005/000983), WO 2005/090354 A1 (PCT/GB2005/000987), and WO 2005/090352 A1 (PCT/EP2005/003038) (all Glaxo Group Limited). PCT/EP2005/003038, PCT/GB2005/000987 and PCT/GB2005/000983, were all filed 15 Mar. 2005.

WO 03/087064 is directed to compounds having both antagonism of the M3 muscarinic receptor and inhibition of PDE4, having the formula:

wherein, inter alia, Y is —NH—R2 or

Two subsequent papers describe in vitro profiles of the lead compounds and in vivo activity after intranasal dosing. Provins, L., et al., Bioorganic & Medicinal Chemistry Letters, 16: 1834-1839 (2006), and Provins, L. et al., Bioorganic & Medicinal Chemistry Letters, 17:3007-3080 (2007). Although promising the data demonstrates that the compounds do not display the in vitro profile that will deliver an in vivo profile displayed by compounds optimized for each molecular target.

Therefore, there is still a need for compounds which contain both the strength and benefit of a combination of PDE4 inhibitory activity and the muscarinic antagonist activity for the treatment of and/or prophylaxis of respiratory diseases, such as chronic obstructive pulmonary disease (COPD), asthma, or inflammatory or allergic diseases such as rhinitis (e.g. allergic rhinitis), atopic dermatitis or psoriasis. The present invention is directed to the novel concept of providing a dual pharmacophore which has both activities.

SUMMARY OF THE INVENTION

The present invention provides for the novel compounds of Formula (I), and pharmaceutical compositions comprising a compound of Formula (I) and a pharmaceutically acceptable carrier or diluent.

Compounds of formula (I) are represented by the structure:

wherein

-   X₁ is oxygen, or N(R_(4a)); -   R_(4a) is hydrogen, or C₁₋₂alkyl; -   R_(5a) is hydrogen, or C₁₋₂alkyl; -   Z is selected from the group consisting of C₁-C₆-alkyl, C(O), S(O)q,     C(O)NH, and C(O)O; -   Z1 is selected from the group consisting of C(O), S(O)q, HNC(O), and     OC(O); -   q is 0 or an integer having a value of 1 or 2; -   v is an integer having a value of 1 to 5; -   n is an integer having a value of 1, 2 or 3; -   n₃ is an integer having a value of 1 to 4; -   Y₄ is independently selected at each occurrence from hydrogen,     halogen, C₁₋₄ alkyl or C₁₋₄ alkoxy, or two of the Y₄ moieties     together with the carbons to which they are attached form a 5-6     membered saturated, partially unsaturated or fully unsaturated C5-C6     ring -   Re and Rf are independently selected at each occurrence from     hydrogen, or C₁₋₄alkyl; -   R¹ is selected from the group consisting of C₁₋₃alkyl,     —CH₂—C₁₋₂-fluoroalkyl, and —CH₂CH₂OH; -   R² is selected from the group consisting of a hydrogen atom,     C₁₋₄alkyl, C₁₋₂-fluoroalkyl, cyclopropyl, cyclobutyl, and     (cyclopropyl)methyl-; -   R³ is selected from the group consisting of an optionally     substituted C₄₋₇cycloalkyl, an optionally substituted     mono-unsaturated-C₅₋₇cycloalkenyl, an optionally substituted     heterocyclic group of sub-formula (aa), (bb) or (cc), and a bicyclic     group of sub-formula (dd), and (ee);

-   -   n¹ is an integer having a value of 1 or 2;     -   n² is an integer having a value of 1 or 2;     -   Y is O, S, SO₂, or NR^(10a);     -   R^(10a) is a hydrogen atom (H), methyl, C(O)NH₂, C(O)-methyl, or         C(O)—C₁fluoroalkyl;     -   Y¹, Y² and Y³ are independently CH₂ or oxygen, provided that no         more than one of Y¹, Y² and Y³ are oxygen;

-   and wherein when R³ is C₄₋₇cycloalkyl it is optionally substituted     on a ring carbon with one or two substituents independently selected     from oxo (═O); OH; methoxy; C₁fluoroalkoxy; NH₂; C₁₋₂alkyl;     C₁fluoroalkyl; —CH₂OH; —CH(Me)OH; —CH₂CH₂OH; —CH₂NH₂; —C(O)OH;     —C(O)NHR²⁴ wherein R²⁴ is H or methyl; —C(O)methyl; fluoro;     hydroxyimino (═N—OH); or (C₁₋₂alkoxy)imino (═N—OR²⁶ where R²⁶ is     C₁₋₂alkyl);     -   and wherein any OH, methoxy, fluoroalkoxy or NH₂ substituent is         not bonded to the R³ ring carbon bonded to the —NH— group of         formula (I); and any OH, methoxy, fluoroalkoxy, —CH₂OH,         —CH(Me)OH, —CH₂CH₂OH, —CH₂NH₂, or —C(O)OH substituent on a ring         carbon of the C₄₋₇cycloalkyl is at the 3-position of a R³         cyclobutyl ring; or at the 3- or 4-position of a R³ cyclopentyl         ring; or at the 3-, 4- or 5-position of a R³ cyclohexyl ring; or         at the 3-, 4-, 5- or 6-position of a R³ cycloheptyl ring;     -   and if the C₄₋₇cycloalkyl is substituted by —C(O)NHR²⁴ or         —C(O)methyl substituent on a ring carbon it is at the 3-position         of the R³ cyclobutyl ring; or at the 3- or 4-position of the R³         cyclopentyl ring; or at the 4-position of a R³ cyclohexyl ring;         or at the 3-, 4-, 5- or 6-position of a R³ cycloheptyl ring         (wherein, in this connection, the 1-position of the R³         cycloalkyl ring is deemed to be the connection point to the —NH—         in formula (I), that is the ring atom connecting to the —NH— in         formula (I));

-   and wherein, when R³ is the optionally substituted heterocyclic     group of sub-formula (aa), (bb) or (cc), then R³ is the heterocyclic     group of sub-formula (aa), (bb) or (cc) optionally substituted on a     ring carbon with one or two oxo (═O) substituents;

-   and wherein, when R³ is optionally substituted     mono-unsaturated-C₅₋₇cycloalkenyl, then the cycloalkenyl is     optionally substituted on a ring carbon with one substituent being     fluoro or methyl, and the R³ ring carbon bonded to the —NH— group of     formula (I) does not partake in the cycloalkenyl double bond;

-   Ar₁ and Ar₂, are independently, selected from the group consisting     of an optionally substituted phenyl and an optionally substituted     monocyclic heteroaryl;

-   R₆ is NR₇R₈, or is a heterocyclic group of the subformula (ff),     (gg), (hh), (ii), (jj), (kk), (ll), (mm) or (nn):

-    or R₆ is an optionally substituted C5-C7 membered ring containing     one or two nitrogens, or a corresponding bicyclic ring containing     one or two nitrogens; or morpholino, 1-azabicyclo[2.2.2]oct-3-yl; or     8-methyl-8-azabicyclo[3.2.1]oct-3-yl; -   R₉ is selected from the group consisting of hydrogen, optionally     substituted C₁₋₆ alkyl, optionally substituted aryl, optionally     substituted arylC₁₋₂alkyl, optionally substituted heteroaryl,     optionally substituted heteroaryl C₁₋₂alkyl, optionally substituted     heterocyclic, optionally substituted heterocyclic C₁₋₂alkyl, and     C(O)C₁₋₂alkyl; -   R_(9a) is selected from the group consisting of hydrogen, optionally     substituted C₁₋₆ alkyl, optionally substituted aryl, optionally     substituted arylC₁₋₂alkyl, optionally substituted heteroaryl,     optionally substituted heteroaryl C₁₋₂alkyl, optionally substituted     heterocyclic, optionally substituted heterocyclic C₁₋₂alkyl, and     C(O)C₁₋₂alkyl; -   Rd is independently selected at each occurrence from the group     consisting of hydrogen, hydroxy, optionally substituted C₁₋₆ alkyl,     amino, optionally substituted aryl, optionally substituted     arylC₁₋₂alkyl, optionally substituted heterocyclic, optionally     substituted heterocyclicC₁₋₂alkyl, optionally substituted     heteroaryl, optionally substituted heteroaryl C₁₋₂alkyl, ═O,     C(O)C₁₋₂alkyl, OC(O)R₁₇, and C(O)N(R₁₀)₂; -   R₁₅ and R₁₆ are independently selected at each occurrence from     hydrogen, or C₁₋₄ alkyl; -   R₁₇ is selected at each occurrence from the group consisting of     optionally substituted C₁₋₄ alkyl, optionally substituted C3-7     cycloalkyl, optionally substituted C₃₋₇ cycloalkylC₁₋₄alkyl,     optionally substituted aryl, optionally substituted arylC₁₋₄alkyl,     heterocyclic, optionally substituted heterocyclic, optionally     substituted heterocyclicC₁₋₄alkyl, optionally substituted     heteroaryl, and optionally substituted heteroaryl C₁₋₄alkyl; -   R_(a) is independently selected at each occurrence from the group     consisting of hydrogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkyl-C₁₋₄alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆C₁₋₄-alkyl, S(O)_(q)C₁₋₄     alkyl, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, C(O)N(R₁₀)₂, optionally     substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally     substituted heterocyclic, optionally substituted heterocyclic C₁₋₄     alkyl, optionally substituted heteroaryl, and optionally substituted     heteroarylC₁₋₄ alkyl; -   R_(a1) is independently selected at each occurrence from the group     consisting of hydrogen, halogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆, NR₁₅R₁₆C₁₋₄alkyl,     S(O)_(q)C₁₋₄ alkyl, hydroxy, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, OC(O)R₁₇,     C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted     arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally     substituted heterocyclic C₁₋₄ alkyl, optionally substituted     heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl; -   R_(b) is independently selected at each occurrence from the group     consisting of hydrogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆C₁₋₄alkyl, S(O)_(q)C₁₋₄     alkyl, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, C(O)N(R₁₀)₂, optionally     substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally     substituted heterocyclic, optionally substituted heterocyclic C₁₋₄     alkyl, optionally substituted heteroaryl, and optionally substituted     heteroarylC₁₋₄ alkyl; -   R_(b1) is independently selected at each occurrence from the group     consisting of hydrogen, halogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆, NR₁₅R₁₆C₁₋₄alkyl,     S(O)_(q)C₁₋₄ alkyl, hydroxy, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, OC(O)R₁₇,     C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted     arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally     substituted heterocyclic C₁₋₄ alkyl, optionally substituted     heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl; -   R_(c) is independently selected at each occurrence from hydrogen or     C₁₋₄ alkyl; -   R₁₀ is independently selected at each occurrence from hydrogen or     C₁₋₄ alkyl; -   R_(13a) is selected from hydrogen, C₁₋₂ alkyl; -   R₁₃ is independently selected from hydrogen, C₁₋₂ alkyl, —CH₂OH,     —CH(CH₃)OH, —CH₂CH₂OH, OH, or ═O; -   X is (C(R₁₃))_(p), or (CR_(e)R_(e))_(s1)—X₂—(CR_(f)R_(f))_(s2); -   X₂ is NR_(13a), O, S(O)m, or C(O); -   m is 0 or an integer having a value of 1, or 2; -   p is an integer having a value of 1 or 2; -   s is 0, or is an integer having a value of 1 or 2; -   s1 is 0 or an integer having a value of 1 to 2; -   s2 is 0 or an integer having a value of 1 to 2, provided that when     R₆ is a heterocyclic group of the subformulas (ff), (ii), (jj) and     (ll), and X₂ is NR_(13a), O, or S(O)m and m is 0 or 1, then s2 is 1     or 2, or X is (CH(R₁₃))p; -   t is an integer having a value of 1 to 4; -   t1 is 0 or an integer having a value of 1 to 4; -   R₄ and R₅ are each independently selected from the group consisting     of hydrogen, optionally substituted C₁₋₄ alkyl, optionally     substituted C₃-C₇ cycloalkyl, optionally substituted C₃-C₇     cycloalkyl C₁₋₄ alkyl, optionally substituted heterocyclic,     optionally substituted heterocyclic C₁₋₄ alkyl, optionally     substituted alkenyl, optionally substituted aryl, optionally     substituted arylC₁₋₄ alkyl optionally substituted heteroaryl, and     optionally substituted heteroaryl C₁₋₄ alkyl; -   R₇ is selected from hydrogen, or an optionally substituted C₁₋₄     alkyl; -   R₈ is (CR_(d1)R_(d1))_(t)—NR₁₁R₁₂ or (CR_(d1)R_(d1))_(t1)—R₁₄; -   R_(d1) is independently at each occurrence selected from the group     consisting of hydrogen, optionally substituted C₁₋₄ alkyl,     optionally substituted aryl, optionally substituted heteroaryl, and     optionally substituted heterocyclic; -   R₁₄ is selected from the group consisting of C₁₋₄ alkyl, C₃-C₆     cycloalkyl, optionally substituted heterocyclic, and optionally     substituted heteroaryl moiety; -   R₁₁ and R₁₂ are independently selected from hydrogen, or C₁₋₄ alkyl;     and the asterix indicating point of attachment of the Z moiety to     the phenyl ring;     or a pharmaceutically acceptable salt thereof.

This invention provides for a method of treating both a muscarinic acetylcholine receptor (mAChR) mediated disease, wherein acetylcholine binds to an M₃ mAChR and a phosphodiesterase type IV (PDE4) mediated disease, whereby the compound also binds to the PDE4 isotype, which comprises administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a mammal in need thereof.

One use of compounds of Formula (I), or pharmaceutically acceptable salt thereof are in the treatment and/or prophylaxis of an inflammatory and/or allergic disease in a mammal.

DETAILED DESCRIPTION OF THE INVENTION

Compounds of the present invention provide for a single compound which has the attributes of each pharmacophore optimized for each molecular target in a balanced fashion. This resulting in vivo profile allows for efficacy and duration of action at both targets, inhibition of PDE4 and antagonism of the mAChR, in a defined dose range. It is now possible to produce a compound which is developable to treat at least two aspects of complex disease etiology, for example bronchoconstriction and inflammation found in diseases such as COPD and asthma.

The present invention is directed to a concept of having dual pharmacophores in one molecule that retains potency across both pharmacological groups. Another aspect of the invention is that in addition to retaining dual pharmacological activity the compounds are developable for commercial activities.

In one embodiment of the invention the compound may be administered to a mammal in needed thereof, suitably one to four times daily, and preferably either a once or a twice daily treatment. Suitably, the compound is administered topically or by inhalation (via nose or mouth) for use in the treatment and/or prophylaxis of a disease for which either pharmacophore has previously been associated with treatment of a PDE4 or an M3 mediated disease. Generally this will be an inflammatory and/or allergic disease, such as the treatment of COPD, asthma, adult respiratory distress syndrome, rhinitis, allergic rhinitis, atopic dermatitis, urticaria, allergic conjunctivitis, psoriasis, ulcerative colitis, or Crohn's disease.

One or more specific compounds within the presently invented compounds may be suitable for use as dual PDE4/mAChR inhibitors via an inhaled route of administration.

One or more specific compounds within the presently invented compounds may be suitable for use as dual PDE4/mAChR inhibitors via an intranasal route of administration.

One or more specific compounds within the presently invented compounds may be suitable for use as dual PDE4/mAChR inhibitors via a topical route of administration.

In compounds of formula (I), R¹ is suitably selected from C₁₋₃alkyl, —CH₂—C₁₋₂fluoroalkyl, or —CH₂CH₂OH. In one embodiment of the invention, R¹ is suitably selected from C₁₋₃alkyl, such as methyl, ethyl, n-propyl, or isopropyl. In another embodiment R¹ is ethyl.

Suitably, R² is hydrogen, C₁₋₄alkyl, such as methyl, ethyl, n-propyl, isopropyl, or n-butyl, a C₁₋₂-fluoroalkyl, cyclopropyl, cyclobutyl, or (cyclopropyl)methyl-. In one embodiment of the invention, R² is methyl, ethyl, n-propyl, isopropyl, or n-butyl. In another embodiment of the invention, R² is ethyl.

Suitably, R³ is optionally substituted C₄₋₇cycloalkyl, or optionally substituted mono-unsaturated-C₅₋₇cycloalkenyl, or an optionally substituted heterocyclic group of sub-formula (aa), (bb) or (cc), or a bicyclic group of sub-formula (dd), or (ee);

Suitably, n¹ and n² are independently selected from an integer having a value of 1 or 2.

Suitably, Y is O, S, SO₂, or NR^(10a). In one embodiment of the invention Y is O.

Suitably, R^(10a) is a hydrogen atom (H), methyl, C(O)NH₂, C(O)-methyl, or C(O)—C₁ fluoroalkyl.

Suitably, Y¹, Y² and Y³ are each independently selected from CH₂ or oxygen, provided that no more than one of Y¹, Y² and Y³ are oxygen.

When R³ is an optionally substituted C₄₋₇cycloalkyl, then the C₄₋₇cycloalkyl ring is optionally substituted on a ring carbon with one or two substituents independently selected from oxo (═O); OH; methoxy; C₁fluoroalkoxy; NH₂; C₁₋₂alkyl; C₁fluoroalkyl; —CH₂OH; —CH(Me)OH; —CH₂CH₂OH; —CH₂NH₂; —C(O)OH; —C(O)NHR²⁴ wherein R²⁴ is H or methyl; —C(O)methyl; fluoro; hydroxyimino (═N—OH); or (C₁₋₂alkoxy)imino (═N—OR²⁶ where R²⁶ is C₁₋₂alkyl); and wherein any OH, methoxy, fluoroalkoxy or NH₂ substituent is not bonded to the R³ ring carbon bonded to the —NH— group of formula (I).

When R³ is the optionally substituted heterocyclic group of sub-formula (aa), (bb) or (cc), then R³ is the heterocyclic group of sub-formula (aa), (bb) or (cc) optionally substituted on a ring carbon with one or two oxo (═O) substituents.

When R³ is optionally substituted mono-unsaturated-C₅₋₇cycloalkenyl, then the cycloalkenyl is optionally substituted on a ring carbon with one substituent being fluoro or methyl, and the R³ ring carbon bonded to the —NH— group of formula (I) does not partake in the cycloalkenyl double bond.

In one embodiment of the invention when R³ is the heterocyclic group of sub-formula (aa) and Y is NR¹⁰, then R¹⁰ is not C(O)-methyl, or C(O)—C₁fluoroalkyl; and when R³ is the heterocyclic group of sub-formula (bb) and Y is NR¹⁰, then R¹⁰ is not methyl; and when R³ is the heterocyclic group of sub-formula (cc), then Y is O, S, SO₂ or NR¹⁰ wherein R¹⁰ is H or methyl.

When R³ is optionally substituted C₄₋₇cycloalkyl, then any —C(O)NHR²⁴ or —C(O)R²⁵ substituent on a ring carbon is: at the 3-position of a R³ cyclobutyl ring; or at the 3- or 4-position of a R³ cyclopentyl ring; or at the 4-position of a R³ cyclohexyl ring; or at the 3-, 4-, 5- or 6-position of a R³ cycloheptyl ring (wherein, in this connection, the 1-position of the R³ cycloalkyl ring is deemed to be the connection point to the —NH— in formula (I), that is the ring atom connecting to the —NH— in formula (I)).

When R³ is optionally substituted C₄₋₇cycloalkyl, then any OH, methoxy, fluoroalkoxy, —CH₂OH, —CH(Me)OH, —CH₂CH₂OH, —CH₂NH₂, or —C(O)OH substituent on a ring carbon is: at the 3-position of a R³ cyclobutyl ring; or at the 3- or 4-position of a R³ cyclopentyl ring; or at the 3-, 4- or 5-position of a R³ cyclohexyl ring; or at the 3-, 4-, 5- or 6-position of a R³ cycloheptyl ring.

In one embodiment of the invention, R³ is the sub-formula (bb) and (cc). In another embodiment of the invention R³ is the sub-formula (bb) and (cc), and n1 and n² independently are 1 or 2. In another embodiment, Y is O, and n1 and n² are 1.

In one embodiment of the invention, R³ is the sub-formula (bb). In another embodiment R³ is the sub-formula (bb), and Y is O. In yet another embodiment R³ is the sub-formula (bb), Y is O, and n¹ is 1.

Suitably, X₁ is oxygen, or N(R_(4a)). In one embodiment of the invention X is N(R_(4a)).

Suitably, R_(4a) is hydrogen, methyl or ethyl. In one embodiment of the invention R_(4a) is hydrogen or methyl. In another embodiment of the invention R_(4a) is hydrogen.

Suitably, R_(5a) is hydrogen, methyl or ethyl. In one embodiment of the invention R_(5a) is hydrogen.

Suitably, v is an integer having a value of 1 to 5. In one embodiment, v is 1. In another embodiment v is 1 and R5a is hydrogen.

Suitably, Y₄ is independently selected at each occurrence from hydrogen, halogen, C₁₋₄ alkyl or C₁₋₄ alkoxy or two of the Y₄ moieties together with the carbons to which they are attached form a 5-6 membered saturated, partially unsaturated or fully unsaturated C5-C6 ring. In one embodiment of the invention, the phenyl ring is substituted one or more times independently with fluorine, chlorine, methyl or methoxy. In another embodiment, two of the Y₄ moieties together with the carbons to which they are attached form a 5-6 membered saturated, partially unsaturated or fully unsaturated C5-C6 ring. In another embodiment the two Y₄ moieties together with the carbons to which they are attached form a C6 unsaturated ring, forming a naphthyl ring. It should be noted that the Y₄ moieties may also be substituted on the newly formed C5-C6 ring.

Suitably, n₃ is an integer having a value of 1 to 4.

Suitably, Z is selected from C(O), S(O)q, C(O)NH or C(O)O.

Suitably, Z1 is selected from C(O), S(O)q, HNC(O), or OC(O).

In one embodiment of the invention, Z and Z1 are both C(O). In another embodiment of the invention Z is C(O) and Z1 is S(O)q. In another embodiment of the invention Z is C(O) and Z1 is HNC(O). In another embodiment of the invention Z is C(O) and Z1 is OC(O). In one embodiment of the invention, Z and Z1 are both S(O)q. In another embodiment of the invention Z is S(O)q, and Z1 is C(O). In another embodiment Z is S(O)q, and Z1 is HNC(O). In another embodiment Z is S(O)q, and Z1 is OC(O). In another embodiment Z is C(O)NH and Z1 is C(O). In another embodiment Z is C(O)NH and Z1 is S(O)q. In another embodiment Z is C(O)NH and Z1 is S(O)q. In another embodiment Z is C(O)NH and Z1 is OC(O). In another embodiment Z is C(O)O and Z1 is HNC(O). In another embodiment Z is C(O)O and Z1 is C(O). In another embodiment Z is C(O)O and Z1 is S(O)q. In another embodiment Z is C(O)O and Z1 is HNC(O). In another embodiment Z is C(O)O and Z1 is OC(O). Suitably, when either Z or Z1 is S(O)q, q is 2.

Suitably, q is independently selected at each occurrence from 0 or an integer having a value of 1 or 2. In one embodiment of the invention q is 2 when Z is S(O)q. In another embodiment of the invention q is 2 when Z1 is S(O)q. In another embodiment of the invention q is 2 when Z and Z1 are both S(O)q.

Suitably, Ar₁ and Ar₂ are independently selected from the group consisting of an optionally substituted phenyl and an optionally substituted monocyclic heteroaryl. In one embodiment, Ar₁ and Ar₂ are independently selected from an optionally substituted aryl. In another embodiment both Ar₁ and Ar₂ are independently selected from an optionally substituted phenyl.

Ar₁ and Ar₂ are independently substituted one or more times, suitably 1 to 4 times, at each occurrence by halogen, such as fluorine, chlorine, bromine or iodine; cyano; hydroxy; hydroxy substituted C₁₋₄alkyl; C₁₋₄ alkoxy, such as methoxy or ethoxy; S(O)_(m′)C₁₋₁₀ alkyl, wherein m′ is 0, 1 or 2, such as methyl thio, methyl sulfinyl or methyl sulfonyl; amino, a mono or di-substituted C₁₋₂alkyl amino; C₁₋₄alkyl, such as methyl, ethyl, propyl, isopropyl, or t-butyl; C₂₋₄alkyl alkenyl, such as ethenyl, 1-propenyl, 2-propenyl, or 2-methyl-1-propenyl; or a halosubstituted C₁₋₄ alkyl, such CH₂F, CH₂CH₂F, or CF₃. In one embodiment of the invention the optional substituents are independently selected from halogen, alkyl, alkoxy, or cyano. In another embodiment the optional substituents are independently selected from fluorine, chlorine, methyl, methoxy or cyano.

Examples of suitable heteroaryl rings for Ar₁ and Ar₂ include, but are not limited to, furyl, pyranyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, oxathiadiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, and triazinyl. In one embodiment of the invention the heteroaryl ring is a pyridine.

In one embodiment of the invention, Ar₁ and Ar₂ are both independently selected from an optionally substituted aryl, preferably an optionally substituted phenyl. In one embodiment of the invention both Ar₁ and Ar₂ are independently selected from optionally substituted phenyls

In one embodiment of the invention, the Ar2 ring is phenyl.

In one embodiment of the invention, the Ar1 ring is a heteroaryl ring. In another embodiment the Ar1 ring is a pyridine ring.

In another embodiment of the invention, the Ar1 ring is a phenyl optionally substituted one or more times independently by halogen, alkyl, alkoxy, or cyano. In another embodiment the Ar1 ring is a phenyl optionally substituted one or more times independently by fluorine, chlorine, methyl, methoxy or cyano.

In another embodiment of the invention, the Ar2 ring is phenyl, and the Ar1 ring is a phenyl optionally substituted one or more times independently by halogen, alkyl, alkoxy, or cyano.

For purposes herein the ring position numbering on the Ar₁ moiety, when Ar₁ is a phenyl ring, is as shown below:

In one embodiment the Ar₁ ring is mono-substituted in the 5- or in the 6-position. In another embodiment if the Ar₁ ring is di-substituted it is substituted in both the 5 and 6-position.

In one embodiment, the Ar₁ 1 ring is an optionally substituted phenyl ring. In another embodiment the phenyl ring is substituted one or more times, suitably 1 to 2 times, by halogen, cyano, or C₁₋₄ alkoxy. In another embodiment, the Ar₁ ring is a phenyl, or an optionally substituted phenyl in the 6-position, such as by fluorine, or methoxy.

Suitably, R₆ is NR₇R₈, or is a heterocyclic group of the subformula (ff), (gg), (hh), (ii), (jj), (kk), (ll), (mm) or (nn):

or R₆ is an optionally substituted C5-C7 membered ring containing one or two nitrogens, or a corresponding bicyclic ring containing one or two nitrogens.

Suitably, when R₆ is an optionally substituted C5-C7 membered ring containing one or two nitrogens, or a corresponding bicyclic ring containing one or two nitrogens the rings include variations of the ring nitrogen positions from subformulas (ff) to (nn). For instance, in formula (ff) the nitrogens are at the 1-4 position, other options include 1-3, or 1-2 nitrogens with similarly substituted Ra, Rb, Rb1, R9, etc. substituents. Some of these exemplified ring systems are shown below:

Suitably s is 0, or is an integer having a value of 1 or 2. In one embodiment of the invention s is 1 or 2. In another embodiment of the invention s is 1.

In one embodiment of the invention, R₆ is a heterocyclic group of the subformula (ff), and s is 1 or 2. In another embodiment, R₆ is a heterocyclic group of the subformula (ff), s is 1 or 2, and Rb is independently selected from hydrogen, or methyl.

In another embodiment, R₆ is a heterocyclic group of the subformula (jj).

Suitably, R₇ is selected from hydrogen, or an optionally substituted C₁₋₄ alkyl. In one embodiment of the invention R₇ is hydrogen or methyl.

Suitably, R₈ is (CR_(d1)R_(d1))_(t)—NR₁₁R₁₂ or (CR_(d1)R_(d1))_(t1)—R₁₄.

Suitably, R_(d1) is independently at each occurrence selected from the group consisting of hydrogen, optionally substituted C₁₋₄ alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic.

Suitably, t is an integer having a value of 1 to 4. In one embodiment, t is 1 or 2.

Suitably, t1 is 0 or an integer having a value of 1 to 4. In one embodiment, t1 is 0, or 1. In another embodiment, t1 is 0.

Suitably, R₁₁ and R₁₂ are independently selected from hydrogen, or C₁₋₄ alkyl.

Suitably, R₁₄ is selected from C₁₋₄ alkyl, C₃-C₆ cycloalkyl, optionally substituted aryl, optionally substituted heterocyclic, or optionally substituted heteroaryl moiety. When R₁₄ is a heterocyclic group of the subformula (ff), (ii), (jj), (ll), (mm) and (nn), then t1 is other than 0.

Suitably, when R₁₄ is a heteroaryl it is a monocyclic five- to seven-membered unsaturated hydrocarbon ring containing at least one heteroatom selected from oxygen, nitrogen and sulfur; or a fused C8-C12 aromatic ring comprising at least one heteroatom selected from oxygen, nitrogen and sulfur.

Examples of heteroaryl rings include, but are not limited to, furyl, pyranyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, oxathiadiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, uracil, indolyl, isoindolyl, indazolyl, indolizinyl, azaindolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, benzothiophenyl, quinolyl, isoquinolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, cinnolinyl, purinyl, and phthalazinyl. In one embodiment when R₁₄ is an optionally substituted heteroaryl it is selected from an optionally substituted thiophenyl, optionally substituted pyridinyl, or an optionally substituted pyrimidinyl.

Suitably, when R₁₄ is a heterocyclic it is a C3-C7 monocyclic non-aromatic hydrocarbon ring containing at least one heteroatom selected from nitrogen, oxygen, sulphur or oxidized sulphur moieties, such as S(O)m, and m is 0 or an integer having a value of 1 or 2, or the heterocyclic is a fused, C8-C12 saturated or partially unsaturated ring system wherein one of the rings may be aromatic, or heteroaromatic. Each of the fused rings may have from four to seven ring atoms. Examples of suitable heterocyclyl groups include, but are not limited to, the saturated or partially saturated versions of the heteroaryl moieties as defined above, such as tetrahydropyrrole, tetrahydropyran, tetrahydrofuran, tetrahydrothiophene (including oxidized versions of the sulfur moiety), azepine, diazepine, aziridinyl, pyrrolinyl, pyrrolidinyl, 2-oxo-1-pyrrolidinyl, 3-oxo-1-pyrrolidinyl, 1,3-benzdioxol-5-yl, imidazolinyl, imidazolidinyl, indolinyl, pyrazolinyl, pyrazolidinyl, piperidinyl, piperazinyl, morpholino and thiomorpholino (including oxidized versions of the sulfur moiety). In one embodiment when R₁₄ is an optionally substituted heterocyclic ring, the ring is an optionally substituted piperidinyl, piperazinyl, optionally substituted oxohexahydro-1H-azepine, or an optionally substituted 3′-[(1-Azabicyclo-[2.2.2]oct-3-yl.

In one embodiment when R₁₄ is a C₃-C₆ cycloalkyl it is suitably selected from cyclopropyl, cyclopentyl, or cyclohexyl. In another embodiment when R₁₄ is a C₁₋₄ alkyl it is ethyl, isopropyl, n-propyl, n-butyl, sec-butyl, or t-butyl.

Suitably, Rd is independently at each occurrence selected from the group consisting of hydrogen, optionally substituted C₁₋₄ alkyl, optionally substituted aryl, optionally substituted heteroaryl, and an optionally substituted heterocyclic moiety. When Rd is an optionally substituted moiety, excluding hydrogen, the moiety may be substituted one or more times, suitably 1 to 4 times, independently by halogen, such as fluorine or chlorine, or a C₁₋₂alkyl. In one embodiment of the invention Rd is independently hydrogen or methyl.

Suitably, R₉ is hydrogen, optionally substituted C₁₋₆ alkyl, optionally substituted aryl, optionally substituted arylC₁₋₂alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C₁₋₂alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic C₁₋₂alkyl, or C(O)C₁₋₂alkyl. When R₉ is an optionally substituted C₁₋₆ alkyl, the alkyl is substituted one or more times, suitably 1 or 2 times independently by halogen, hydroxy, NR₁₅R₁₆, C₁₋₄ alkoxy, S(O)qC₁₋₄ alkyl. In one embodiment of the invention, R₉ is hydrogen or methyl.

Suitably, R_(9a) is hydrogen, optionally substituted C₁₋₆ alkyl, optionally substituted aryl, optionally substituted arylC₁₋₂alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C₁₋₂alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic C₁₋₂alkyl, C(O)C₁₋₂alkyl. In one embodiment of the invention, R_(9a) is hydrogen or optionally substituted C₁₋₃ alkyl.

Suitably, R_(a) is independently selected at each occurrence from hydrogen, C₁₋₄ alkyl, C₃₋₇cycloalkyl, C₃₋₇cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆C₁₋₄alkyl, S(O)_(q)C₁₋₄ alkyl, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, OC(O)C₁₋₄ alkyl, C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic C₁₋₄ alkyl, optionally substituted heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl. In one embodiment, R_(a) is independently hydrogen, or methyl.

Suitably, R_(a1) is independently selected at each occurrence from hydrogen, halogen, optionally substituted C₁₋₄ alkyl, optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇ cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆, NR₁₅R₁₆C₁₋₄alkyl, S(O)_(q)C₁₋₄ alkyl, hydroxy, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, OC(O)R₁₇, C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic C₁₋₄ alkyl, optionally substituted heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl. In one embodiment, R_(a) is independently hydrogen, or methyl.

Suitably, R_(b) is independently selected at each occurrence from hydrogen, optionally substituted C₁₋₄ alkyl, optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆C₁₋₄alkyl, S(O)_(q)C₁₋₄ alkyl, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic C₁₋₄ alkyl, optionally substituted heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl. In one embodiment of the invention, R_(b) is independently selected from hydrogen or methyl.

Suitably, R_(b1) is independently selected at each occurrence from hydrogen, halogen, optionally substituted C₁₋₄ alkyl, optionally substituted C₃₋₇cycloalkyl, optionally substituted C₃₋₇ cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆, NR₁₅R₁₆C₁₋₄alkyl, S(O)_(q)C₁₋₄ alkyl, hydroxy, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, OC(O)R₁₇, C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic C₁₋₄ alkyl, optionally substituted heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl. In one embodiment of the invention, R_(b1) is independently selected from hydrogen or methyl.

Suitably, R_(d) is independently selected at each occurrence from hydrogen, hydroxy, optionally substituted C₁₋₆ alkyl, amino, optionally substituted aryl, optionally substituted arylC₁₋₂alkyl, optionally substituted heterocyclic, optionally substituted heterocyclicC₁₋₂alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C₁₋₂alkyl, ═O, C(O)C₁₋₂alkyl, OC(O)R₁₇, or C(O)N(R₁₀)₂. When R_(d) is an optionally substituted C₁₋₆ alkyl, the alkyl is substituted one or more times, suitably 1 or 2 times independently by halogen, hydroxy, NR₁₅R₁₆, C₁₋₄ alkoxy, S(O)qC₁₋₄ alkyl. In one embodiment of the invention, R_(d) is hydrogen or methyl.

Suitably, R_(c) is independently selected at each occurrence from hydrogen or C₁₋₄ alkyl.

Suitably, R₁₀ is independently selected from hydrogen or C₁₋₄ alkyl.

Suitably, R₁₅ and R₁₆ are independently selected from hydrogen, or C₁₋₄ alkyl. In one embodiment of the invention R₁₅ and R₁₆ are hydrogen or methyl.

Suitably, R₁₇ is selected from optionally substituted C₁₋₄ alkyl, optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇ cycloalkylC₁₋₄alkyl, optionally substituted aryl, optionally substituted arylC₁₋₄alkyl, heterocyclic, optionally substituted heterocyclic, optionally substituted heterocyclicC₁₋₄alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C₁₋₄alkyl.

Suitably, X is (C(R₁₃))p, or (CR_(e)R_(e))_(s1)—X₂—(CR_(f)R_(f))_(s2).

Suitably, X₂ is NR_(13a), O, S(O)m, or C(O).

Suitably, R₁₃ is selected from hydrogen, C₁₋₂ alkyl, —CH₂OH, —CH(CH₃)OH, —CH₂CH₂OH, OH, or ═O. In one embodiment of the invention R₁₃ is hydrogen.

Suitably, R_(13a) is selected from hydrogen, C₁₋₂ alkyl. In one embodiment of the invention R₁₃ is hydrogen.

Suitably, s1 is 0 or an integer having a value of 1 to 2. In one embodiment of the invention s1 is 0.

Suitably, s2 is 0 or an integer having a value of 1 to 2. However, when R₆ is a heterocyclic group of the subformulas (ff), (ii), (jj) and (ll), and X₂ is NR_(13a), O, or S(O)m (and m is 0 or 1) then s2 is 1 or 2, or X is the group (CH(R₁₃))p.

Suitably, p is an integer having a value of 1 or 2.

Suitably, n is an integer having a value of 1, 2 or 3.

Suitably, n₃ is an integer having a value of 1 to 3.

Suitably, m is 0 or an integer having a value of 1, or 2.

Suitably, Y₄ is independently selected at each occurrence from hydrogen, halogen, or C₁₋₄ alkyl; C₃-C₇ cycloalkyl, or C₃-C₇ cycloalkyl C₁₋₄ alkyl. In one embodiment of the invention Y₄ is hydrogen, and n₃ is 1.

Suitably, R₄ and R₅ are each independently selected from the group consisting of hydrogen, optionally substituted C₁₋₄alkyl, optionally substituted C₃-C₇ cycloalkyl, optionally substituted C₃-C₇ cycloalkyl C₁₋₄alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic C₁₋₄alkyl, optionally substituted C₂₋₄ alkenyl, optionally substituted aryl, optionally substituted aryl C₁₋₄alkyl optionally substituted heteroaryl, and optionally substituted heteroaryl C₁₋₄alkyl.

In one embodiment R₅ is hydrogen, and n is 1. In one embodiment R₄ is hydrogen, or C₁₋₄alkyl. In another embodiment, R₄ and R₅ are both hydrogen, and n is 1.

One embodiment of the invention are compounds of formula (Ia), a subset of compounds of Formula (I) shown above. The asterix indicating point of attachment of the Z moiety in the 1-3 or 1-4 position to the phenyl ring on the right hand side of the molecule.

wherein,

-   R¹ is C₁₋₄alkyl; -   R² is a C₁₋₄alkyl; -   Z is (O) or S(O)q; -   Z1 is C(O) or S(O)q; -   Z2 and Z3 are independently at each occurrence selected from the     group consisting of hydrogen, halogen, cyano and C₁₋₄alkoxy; -   v is an integer having a value of 1 to 5; -   R4a is selected from hydrogen or C1-2 alkyl; -   R5a is selected from hydrogen or C1-2 alkyl; -   R₆ is NR₇R₈, or is a heterocyclic group of the subformula (ff),     (gg), (hh), (ii), (jj), (kk), (ll), (mm) or (nn):

-    or R₆ is an optionally substituted C₅-C₇ membered ring containing     one or two nitrogens, or a corresponding bicyclic ring containing     one or two nitrogens; -   R₉ is selected from the group consisting of hydrogen, optionally     substituted C₁₋₆ alkyl, optionally substituted aryl, optionally     substituted arylC₁₋₂alkyl, optionally substituted heteroaryl,     optionally substituted heteroaryl C₁₋₂alkyl, optionally substituted     heterocyclic, optionally substituted heterocyclic C₁₋₂alkyl, and     C(O)C₁₋₂alkyl; -   R_(9a) is selected from the group consisting of hydrogen, optionally     substituted C₁₋₆ alkyl, optionally substituted aryl, optionally     substituted arylC₁₋₂alkyl, optionally substituted heteroaryl,     optionally substituted heteroaryl C₁₋₂alkyl, optionally substituted     heterocyclic, optionally substituted heterocyclic C₁₋₂alkyl, and     C(O)C₁₋₂alkyl; -   Rd is independently selected at each occurrence from the group     consisting of hydrogen, hydroxy, optionally substituted C₁₋₆ alkyl,     amino, optionally substituted aryl, optionally substituted     arylC₁₋₂alkyl, optionally substituted heterocyclic, optionally     substituted heterocyclicC₁₋₂alkyl, optionally substituted     heteroaryl, optionally substituted heteroaryl C₁₋₂alkyl, ═O,     C(O)C₁₋₂alkyl, OC(O)R₁₇, and C(O)N(R₁₀)₂; -   R₁₅ and R₁₆ are independently selected at each occurrence from     hydrogen, or C₁₋₄ alkyl; -   R₁₇ is selected at each occurrence from the group consisting of     optionally substituted C₁₋₄ alkyl, optionally substituted C3-7     cycloalkyl, optionally substituted C₃₋₇ cycloalkylC₁₋₄alkyl,     optionally substituted aryl, optionally substituted arylC₁₋₄alkyl,     heterocyclic, optionally substituted heterocyclic, optionally     substituted heterocyclicC₁₋₄alkyl, optionally substituted     heteroaryl, and optionally substituted heteroaryl C₁₋₄alkyl; -   R_(a) is independently selected at each occurrence from the group     consisting of hydrogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkyl-C₁₋₄alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆C₁₋₄alkyl, S(O)_(q)C₁₋₄     alkyl, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, C(O)N(R₁₀)₂, optionally     substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally     substituted heterocyclic, optionally substituted heterocyclic C₁₋₄     alkyl, optionally substituted heteroaryl, and optionally substituted     heteroarylC₁₋₄ alkyl; -   R_(a1) is independently selected at each occurrence from the group     consisting of hydrogen, halogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆, NR₁₅R₁₆C₁₋₄alkyl,     S(O)_(q)C₁₋₄ alkyl, hydroxy, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, OC(O)R₁₇,     C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted     arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally     substituted heterocyclic C₁₋₄ alkyl, optionally substituted     heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl; -   R_(b) is independently selected at each occurrence from the group     consisting of hydrogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆C₁₋₄alkyl, S(O)_(q)C₁₋₄     alkyl, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, C(O)N(R₁₀)₂, optionally     substituted aryl, optionally substituted arylC₁₋₄ alkyl, optionally     substituted heterocyclic, optionally substituted heterocyclic C₁₋₄     alkyl, optionally substituted heteroaryl, and optionally substituted     heteroarylC₁₋₄ alkyl; -   R_(b1) is independently selected at each occurrence from the group     consisting of hydrogen, halogen, optionally substituted C₁₋₄ alkyl,     optionally substituted C₃₋₇ cycloalkyl, optionally substituted C₃₋₇     cycloalkylC₁₋₄ alkyl, C₁₋₄ alkoxy, NR₁₅R₁₆, NR₁₅R₁₆C₁₋₄alkyl,     S(O)_(q)C₁₋₄ alkyl, hydroxy, ═O, —CH(O), C(O)₂C₁₋₄ alkyl, OC(O)R₁₇,     C(O)N(R₁₀)₂, optionally substituted aryl, optionally substituted     arylC₁₋₄ alkyl, optionally substituted heterocyclic, optionally     substituted heterocyclic C₁₋₄ alkyl, optionally substituted     heteroaryl, and optionally substituted heteroarylC₁₋₄ alkyl; -   R_(c) is independently selected at each occurrence from hydrogen or     C₁₋₄ alkyl; -   R₁₀ is independently selected at each occurrence from hydrogen or     C₁₋₄ alkyl; -   q is 0 or an integer having a value of 1 or 2; -   t is an integer having a value of 1 to 4; -   t1 is 0 or an integer having a value of 1 to 4; -   R₄ and R₅ are each independently selected from the group consisting     of hydrogen, optionally substituted C₁₋₄ alkyl, optionally     substituted C₃-C₇ cycloalkyl, optionally substituted C₃-C₇     cycloalkyl C₁₋₄ alkyl, optionally substituted heterocyclic,     optionally substituted heterocyclic C₁₋₄ alkyl, optionally     substituted alkenyl, optionally substituted aryl, optionally     substituted arylC₁₋₄ alkyl optionally substituted heteroaryl, and     optionally substituted heteroaryl C₁₋₄ alkyl; -   R₇ is selected from hydrogen, or an optionally substituted C₁₋₄     alkyl; -   R₈ is (CR_(d1)R_(d1))_(t)—NR₁₁R₁₂ or (CR_(d1)R_(d1))_(t1)—R₁₄; -   R_(d1) is independently at each occurrence selected from the group     consisting of hydrogen, optionally substituted C₁₋₄ alkyl,     optionally substituted aryl, optionally substituted heteroaryl, and     optionally substituted heterocyclic; -   R₁₄ is selected from the group consisting of C₁₋₄ alkyl, C₃-C₆     cycloalkyl, optionally substituted heterocyclic, and optionally     substituted heteroaryl moiety; -   R₁₁ and R₁₂ are independently selected from hydrogen, or C₁₋₄ alkyl;     and the asterix indicating point of attachment of the Z moiety to     the phenyl ring; or a pharmaceutically acceptable salt thereof.

Compounds of Formula (Ia) wherein one of Z or Z1 is a S(O)q, and q=2 are exemplified by compounds of Examples 329 to 335.

For purposes herein, all substituents for Formula (Ia) are as defined above for Formula (I).

It is to be understood that the present invention covers all combinations of particular and preferred groups described hereinabove. It is also to be understood that the present invention encompasses compounds in which a particular group or parameter, e.g. S(O)m, etc. may occur more than once. In such compounds it will be appreciated that each group or parameter is independently selected from the values listed. When any variable occurs more than one time in a formula, its definition on each occurrence is independent of its definition at every other occurrence.

Particular compounds according to the invention include those mentioned in the examples and their pharmaceutically acceptable derivatives.

As used herein, the term “pharmaceutically acceptable” means a compound which is suitable for pharmaceutical and veterinary usage. Salts and solvates of compounds of the invention which are suitable for use in medicine are those wherein the counter-ion or associated solvent is pharmaceutically acceptable. However, salts and solvates having non-pharmaceutically acceptable counter-ions or associated solvents are within the scope of the present invention, for example, for use as intermediates in the preparation of other compounds of the invention and their pharmaceutically acceptable salts and solvates.

As used herein, the term “pharmaceutically acceptable derivative”, means any pharmaceutically acceptable salt, solvate or prodrug e.g. ester, of a compound of the invention, which upon administration to the recipient is capable of providing (directly or indirectly) a compound of the invention, or an active metabolite or residue thereof. Such derivatives are recognizable to those skilled in the art, without undue experimentation. Nevertheless, reference is made to the teaching of Burger's Medicinal Chemistry and Drug Discovery, 5^(th) Edition, Vol. 1: Principles and Practice, which is incorporated herein by reference to the extent of teaching such derivatives. In one embodiment pharmaceutically acceptable derivatives are salts, solvates, esters, carbamates and phosphate esters. In another embodiment pharmaceutically acceptable derivatives are salts, solvates and esters. In yet another embodiment of the invention pharmaceutically acceptable derivatives are salts and esters, in particular salts.

The compounds of the present invention may be in the form of and/or may be administered as a pharmaceutically acceptable salt. For a review on suitable salts see Berge et al., J. Pharm. Sci., 1977, 66, 1-19.

Typically, a pharmaceutical acceptable salt may be readily prepared by using a desired acid or base as appropriate. The salt may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent.

Salts of the compounds of the present invention may, for example, comprise acid addition salts resulting from reaction of an acid with a nitrogen atom present in a compound of formula (I). Salts encompassed within the term “pharmaceutically acceptable salts” refer to non-toxic salts of the compounds of this invention. Suitable addition salts are formed from acids which form non-toxic salts and examples are acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, ethanesulphonate, formate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydrogen phosphate, hydroiodide, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, monopotassium maleate, mucate, napsylate, nitrate, N-methylglucamine, oxalate, oxaloacetate, pamoate (embonate), palmitate, pantothenate, phosphate/diphosphate, piruvate, polygalacturonate, saccharate, salicylate, stearate, subacetate, succinate, sulphate, tannate, tartrate, teoclate, tosylate, triethiodide, trifluoroacetate and valerate.

Pharmaceutically acceptable base salts include ammonium salts such as a trimethylammonium salt, alkali metal salts such as those of sodium and potassium, alkaline earth metal salts such as those of calcium and magnesium and salts with organic bases, including salts of primary, secondary and tertiary amines, such as isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexyl amine and N-methyl-D-glucamine.

Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as “solvates”. As used herein, the term “solvate” refers to a complex of variable stoichiometry formed by a solute (in this invention, a compound of Formula (I), or a salt thereof) and a solvent. Such solvents for the purpose of the invention may not interfere with the biological activity of the solute. Examples of suitable solvents include water, methanol, ethanol and acetic acid. Preferably the solvent used is a pharmaceutically acceptable solvent. Examples of suitable pharmaceutically acceptable solvents include water, ethanol and acetic acid. Most preferably the solvent used is water. A complex with water is known as a “hydrate”. Solvates of the compound of the invention are within the scope of the invention.

As used herein, the term “prodrug” means a compound which is converted within the body, e.g. by hydrolysis in the blood, into its active form that has medical effects. Pharmaceutically acceptable prodrugs are described in T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series; Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987; and in D. Fleisher, S. Ramon and H. Barbra “Improved oral drug delivery: solubility limitations overcome by the use of prodrugs”, Advanced Drug Delivery Reviews (1996) 19(2) 115-130, each of which are incorporated herein by reference.

Prodrugs are any covalently bonded carriers that release a compound of formula (I) in vivo when such prodrug is administered to a patient. Prodrugs are generally prepared by modifying functional groups in a way such that the modification is cleaved, either by routine manipulation or in vivo, yielding the parent compound. Prodrugs include, for example, compounds of this invention wherein hydroxy or amine groups are bonded to any group that, when administered to a patient, cleaves to form the hydroxy or amine groups. Thus, representative examples of prodrugs include (but are not limited to) acetate, formate and benzoate derivatives of alcohol and amine functional groups of the compounds of formula (I). Further, in the case of a carboxylic acid (—COOH), esters may be employed, such as methyl esters, ethyl esters, and the like. Esters may be active in their own right and/or be hydrolysable under in vivo conditions in the human body. Suitable pharmaceutically acceptable in vivo hydrolysable ester groups include those which break down readily in the human body to leave the parent acid or its salt.

As used herein, “optionally substituted” unless specifically defined shall mean such groups as halogen, such as fluorine, chlorine, bromine or iodine; hydroxy; hydroxy substituted C₁₋₁₀alkyl; C₁₋₁₀ alkoxy, such as methoxy or ethoxy; halosubstituted C₁₋₁₀ alkoxy; S(O)m alkyl, such as methyl thio, methylsulfinyl or methyl sulfonyl; a ketone (—C(O)), or an aldehyde (—C(O)R_(6′)), such as C(O)C₁₋₁₀ alkyl or C(O)aryl, wherein R_(6′) is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, heterocyclyl, heterocyclyl C₁₋₁₀alkyl, aryl, arylC₁₋₁₀ alkyl, heteroaryl or heteroarylC₁₋₁₀ alkyl (and wherein the R_(6′) moieties, excluding hydrogen, may themselves be optionally substituted 1 or 2 times, independently by halogen; hydroxy; hydroxy substituted alkyl; C₁₋₄ alkoxy; S(O)_(m)C₁₋₄ alkyl; amino, mono & di-substituted C₁₋₄ alkyl amino; C₁₋₄ alkyl, or CF₃); C(O)OR_(6′); NR_(4′)R_(14′), wherein R_(4′) and R_(14′) are each independently hydrogen or C₁₋₄ alkyl, such as amino or mono or -disubstituted C₁₋₄ alkyl or wherein the R_(4′)R_(14′) can cyclize together with the nitrogen to which they are attached to form a 5 to 7 membered ring which optionally contains an additional heteroatom selected from O/N/S; C₁₋₁₀ alkyl, C₃₋₇cycloalkyl, or C₃₋₇cycloalkyl C₁₋₁₀ alkyl group, such as methyl, ethyl, propyl, isopropyl, t-butyl, etc. or cyclopropyl methyl; halosubstituted C₁₋₁₀ alkyl, such CF₂CF₂H, or CF₃; an optionally substituted aryl, such as phenyl, or an optionally substituted arylalkyl, such as benzyl or phenethyl, wherein these aryl containing moieties may also be substituted one to two times by halogen; hydroxy; hydroxy substituted alkyl; C₁₋₄ alkoxy; S(O)_(m) C₁₋₄ alkyl; amino, mono & di-substituted C₁₋₄ alkyl amino; C₁₋₄ alkyl, or CF₃.

The term “halo” or “halogens” is used herein to mean the halogens, chloro, fluoro, bromo and iodo.

As used herein, the term “C₁₋₁₀alkyl” or “alkyl” or “alkyl₁₋₁₀” is used herein to mean both straight and branched hydrocarbon chain containing the specified number of carbon atoms, e.g. C₁₋₁₀alkyl means a straight of branched alkyl chain of at least 1, and at most 10, carbon atoms, unless the chain length is otherwise limited. Examples of “alkyl” as used herein include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, isobutyl, isopropyl, sec-butyl, tert-butyl or t-butyl and hexyl and the like.

As used herein, the term “alkenyl” refers to straight or branched hydrocarbon chains containing the specified number of carbon atoms and containing at least one double bond. For example, C₂₋₆alkenyl means a straight or branched alkenyl containing at least 2, and at most 6, carbon atoms and containing at least one double bond. Examples of “alkenyl” as used herein include, but are not limited to ethenyl, 2-propenyl, 3-butenyl, 2-butenyl, 2-pentenyl, 3-pentenyl, 3-methyl-2-butenyl, 3-methylbut-2-enyl, 3-hexenyl, 1,1-dimethylbut-2-enyl and the like.

As used herein, the term “alkoxy” refers to straight or branched chain alkoxy groups containing the specified number of carbon atoms. For example, C₁₋₆alkoxy means a straight or branched alkoxy containing at least 1, and at most 6, carbon atoms. Examples of “alkoxy” as used herein include, but are not limited to, methoxy, ethoxy, propoxy, prop-2-oxy, butoxy, but-2-oxy, 2-methylprop-1-oxy, 2-methylprop-2-oxy, pentoxy and hexyloxy.

As used herein, the term “cycloalkyl” refers to cyclic radicals, such as a non-aromatic hydrocarbon ring containing a specified number of carbon atoms. For example, C₃₋₇cycloalkyl means a non-aromatic ring containing at least three, and at most seven, ring carbon atoms. Representative examples of “cycloalkyl” as used herein include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl and the like.

The term “cycloalkenyl” is used herein to mean cyclic radicals, such as a non-aromatic hydrocarbon ring containing a specified number of carbon atoms preferably of 5 to 7 carbons, which have at least one bond including but not limited to cyclopentenyl, cyclohexenyl, and the like.

The term “alkenyl” is used herein at all occurrences to mean straight or branched chain radical of 2-10 carbon atoms, unless the chain length is limited thereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl and the like.

The term “aryl” is used herein to mean phenyl, naphthyl, and indene.

The terms “heteroaryl ring”, “heteroaryl moiety”, and “heteroaryl” are used herein to mean a monocyclic five- to seven-membered unsaturated hydrocarbon ring containing at least one heteroatom selected from oxygen, nitrogen and sulfur. Examples of heteroaryl rings include, but are not limited to, furyl, pyranyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, oxathiadiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and uracil. The terms “heteroaryl ring”, “heteroaryl moiety”, and “heteroaryl” shall also used herein to refer to fused aromatic rings comprising at least one heteroatom selected from oxygen, nitrogen and sulfur. Each of the fused rings may contain five or six ring atoms. Examples of fused aromatic rings include, but are not limited to, indolyl, isoindolyl, indazolyl, indolizinyl, azaindolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, benzothiophenyl, quinolyl, isoquinolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, cinnolinyl, purinyl, and phthalazinyl.

The terms “heterocyclic rings”, “heterocyclic moieties”, and “heterocyclyl” is used herein to mean a monocyclic three- to seven-membered saturated or non-aromatic, unsaturated hydrocarbon ring containing at least one heteroatom selected from nitrogen, oxygen, sulphur or oxidized sulphur moieties, such as S(O)m, and m is 0 or an integer having a value of 1 or 2. The terms “heterocyclic rings”, “heterocyclic moieties”, and “heterocyclyl” shall also refer to fused rings, saturated or partially unsaturated, and wherein one of the rings may be aromatic, or heteroaromatic. Each of the fused rings may have from four to seven ring atoms. Examples of heterocyclyl groups include, but are not limited to, the saturated or partially saturated versions of the heteroaryl moieties as defined above, such as tetrahydropyrrole, tetrahydropyran, tetrahydrofuran, tetrahydrothiophene (including oxidized versions of the sulfur moiety), azepine, diazepine, aziridinyl, pyrrolinyl, pyrrolidinyl, 2-oxo-1-pyrrolidinyl, 3-oxo-1-pyrrolidinyl, 1,3-benzdioxol-5-yl, imidazolinyl, imidazolidinyl, indolinyl, pyrazolinyl, pyrazolidinyl, piperidinyl, piperazinyl, morpholino and thiomorpholino (including oxidized versions of the sulfur moiety).

The term “arylalkyl” or “heteroarylalkyl”, “heterocyclicalkyl” or “cycloalkylalkyl” as used herein means a C₁₋₄ alkyl (as defined above) attached to an aryl, heteroaryl, heterocyclic or cycloalkyl moiety (as also defined above) unless otherwise indicated.

The term “sulfinyl” is used herein to mean the oxide S(O) of the corresponding sulfide, the term “thio” refers to the sulfide, and the term “sulfonyl” refers to the fully oxidized S(O)₂ moiety.

The term “aroyl” is used herein to mean C(O)Ar, wherein Ar is as phenyl, naphthyl, or aryl alkyl derivative such as defined above, such group include but are not limited to benzyl and phenethyl.

The term “alkanoyl” is used herein to mean C(O)C₁₋₁₀ alkyl wherein the alkyl is as defined above.

As used herein, the term “optionally” means that the subsequently described event(s) may or may not occur, and includes both event(s) which occur and events that do not occur.

As used herein, the term “substituted” refers to substitution with the named substituent or substituents, multiple degrees of substitution being allowed unless otherwise stated.

With regard to stereoisomers, the compounds of the Formulas herein may have one or more asymmetric carbon atom and may occur as racemates, racemic mixtures and as individual enantiomers or diastereomers. All such isomeric forms are included within the present invention, including mixtures thereof.

C is (E) and trans (Z) isomerism may also occur. The present invention includes the individual stereoisomers of the compound of the invention and where appropriate, the individual tautomeric forms thereof, together with mixtures thereof.

Separation of diastereoisomers or cis and trans isomers may be achieved by conventional techniques, e.g. by fractional crystallisation, chromatography or H.P.L.C. A stereoisomeric mixture of the agent may also be prepared from a corresponding optically pure intermediate or by resolution, such as H.P.L.C. of the corresponding racemate using a suitable chiral support or by fractional crystallisation of the diastereoisomeric salts formed by reaction of the corresponding racemate with a suitable optically active acid or base, as appropriate.

Furthermore, some of the crystalline forms of the compounds of the Formulas herein may exist as polymorphs, which are included in the present invention.

Exemplified compounds of the compounds of this invention include the racemates, or optically active forms of the compounds of the working examples herein, and pharmaceutically acceptable salts thereof.

The compounds of this invention may be made by a variety of methods, including standard chemistry. Any previously defined variable will continue to have the previously defined meaning unless otherwise indicated. Illustrative general synthetic methods are set out below and then specific compounds of the invention are prepared in the working Examples.

Methods of Treatment

In order to use a compound of Formula (I) or a pharmaceutically acceptable salt thereof in therapy, it will normally be formulated into a pharmaceutical composition in accordance with standard pharmaceutical practice. This invention, therefore, also relates to a pharmaceutical composition comprising an effective amount of a compound of Formula (I) and a pharmaceutically acceptable carrier or diluent.

Compounds of Formula (I), pharmaceutically acceptable salts thereof and pharmaceutical compositions incorporating such may conveniently be administered by any of the routes conventionally used for drug administration, for instance, orally, topically, parenterally or by inhalation. The compounds of Formula (I) may be administered in conventional dosage forms prepared by combining a compound of Formula (I) with standard pharmaceutical carriers according to conventional procedures. The compounds of Formula (I) may also be administered in conventional dosages in combination with a known, second therapeutically active compound. These procedures may involve mixing, granulating and compressing or dissolving the ingredients as appropriate to the desired preparation. It will be appreciated that the form and character of the pharmaceutically acceptable character or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. The carrier(s) must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The pharmaceutical carrier employed may be, for example, either a solid or liquid. Exemplary of solid carriers are lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary of liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly, the carrier or diluent may include time delay material well known to the art, such as glyceryl mono-stearate or glyceryl distearate alone or with a wax.

A wide variety of pharmaceutical forms can be employed. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form of a troche or lozenge. The amount of solid carrier will vary widely but preferably will be from about 25 mg. to about 1 g. When a liquid carrier is used, the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampule or nonaqueous liquid suspension.

Compounds of Formula (I) may be administered topically, that is by non-systemic administration. This includes the application of a compound of Formula (I) externally to the epidermis or the buccal cavity and the instillation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.

Compounds of Formula (I) may be administered parenterally, that is by intravenous, intramuscular, subcutaneous intranasal, intrarectal, intravaginal or intraperitoneal administration. The subcutaneous and intramuscular forms of parenteral administration are generally preferred. Appropriate dosage forms for such administration may be prepared by conventional techniques. Compounds of Formula (I) may also be administered by inhalation, that is by intranasal and oral inhalation administration. Appropriate dosage forms for such administration, such as an aerosol formulation or a metered dose inhaler, may be prepared by conventional techniques.

In one embodiment of the present invention, the agents of the present invention are delivered via oral inhalation or intranasal administration. Appropriate dosage forms for such administration, such as an aerosol formulation or a metered dose inhaler, may be prepared by conventional techniques.

For administration by inhalation the compounds may be delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane such as tetrafluoroethane or heptafluoropropane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention and a suitable powder base such as lactose or starch.

Dry powder compositions for topical delivery to the lung by inhalation may, for example, be presented in capsules and cartridges of for example gelatine or blisters of for example laminated aluminum foil, for use in an inhaler or insufflator. Powder blend formulations generally contain a powder mix for inhalation of the compound of the invention and a suitable powder base (carrier/diluent/excipient substance) such as mono-, di- or poly-saccharides (e.g. lactose or starch).

Each capsule or cartridge may generally contain between 20 μg-10 mg of the compound of formula (I) optionally in combination with another therapeutically active ingredient. Alternatively, the compound of the invention may be presented without excipients.

Suitably, the packing/medicament dispenser is of a type selected from the group consisting of a reservoir dry powder inhaler (RDPI), a multi-dose dry powder inhaler (MDPI), and a metered dose inhaler (MDI).

By reservoir dry powder inhaler (RDPI) it is meant an inhaler having a reservoir form pack suitable for comprising multiple (un-metered doses) of medicament in dry powder form and including means for metering medicament dose from the reservoir to a delivery position. The metering means may for example comprise a metering cup, which is movable from a first position where the cup may be filled with medicament from the reservoir to a second position where the metered medicament dose is made available to the patient for inhalation.

By multi-dose dry powder inhaler (MDPI) is meant an inhaler suitable for dispensing medicament in dry powder form, wherein the medicament is comprised within a multi-dose pack containing (or otherwise carrying) multiple, define doses (or parts thereof) of medicament. In a preferred aspect, the carrier has a blister pack form, but it could also, for example, comprise a capsule-based pack form or a carrier onto which medicament has been applied by any suitable process including printing, painting and vacuum occlusion.

In the case of multi-dose delivery, the formulation can be pre-metered (e.g. as in Diskus, see GB 2242134, U.S. Pat. Nos. 6,632,666, 5,860,419, 5,873,360 and 5,590,645 or Diskhaler, see GB 2178965, 2129691 and 2169265, U.S. Pat. Nos. 4,778,054, 4,811,731, 5,035,237, the disclosures of which are hereby incorporated by reference) or metered in use (e.g. as in Turbuhaler, see EP 69715 or in the devices described in U.S. Pat. No. 6,321,747 the disclosures of which are hereby incorporated by reference). An example of a unit-dose device is Rotahaler (see GB 2064336 and U.S. Pat. No. 4,353,656, the disclosures of which are hereby incorporated by reference).

The Diskus inhalation device comprises an elongate strip formed from a base sheet having a plurality of recesses spaced along its length and a lid sheet hermetically but peelably sealed thereto to define a plurality of containers, each container having therein an inhalable formulation containing a compound of Formula (I) preferably combined with lactose. Preferably, the strip is sufficiently flexible to be wound into a roll. The lid sheet and base sheet will preferably have leading end portions which are not sealed to one another and at least one of the said leading end portions is constructed to be attached to a winding means. Also, preferably the hermetic seal between the base and lid sheets extends over their whole width. The lid sheet may preferably be peeled from the base sheet in a longitudinal direction from a first end of the said base sheet.

In one aspect, the multi-dose pack is a blister pack comprising multiple blisters for containment of medicament in dry powder form. The blisters are typically arranged in regular fashion for ease of release of medicament there from.

In one aspect, the multi-dose blister pack comprises plural blisters arranged in generally circular fashion on a disc-form blister pack. In another aspect, the multi-dose blister pack is elongate in form, for example comprising a strip or a tape.

In one aspect, the multi-dose blister pack is defined between two members peelably secured to one another. U.S. Pat. Nos. 5,860,419; 5,873,360 and 5,590,645 describe medicament packs of this general type. In this aspect, the device is usually provided with an opening station comprising peeling means for peeling the members apart to access each medicament dose. Suitably, the device is adapted for use where the peelable members are elongate sheets which define a plurality of medicament containers spaced along the length thereof, the device being provided with indexing means for indexing each container in turn. More preferably, the device is adapted for use where one of the sheets is a base sheet having a plurality of pockets therein, and the other of the sheets is a lid sheet, each pocket and the adjacent part of the lid sheet defining a respective one of the containers, the device comprising driving means for pulling the lid sheet and base sheet apart at the opening station.

By metered dose inhaler (MDI) it is meant a medicament dispenser suitable for dispensing medicament in aerosol form, wherein the medicament is comprised in an aerosol container suitable for containing a propellant-based aerosol medicament formulation. The aerosol container is typically provided with a metering valve, for example a slide valve, for release of the aerosol form medicament formulation to the patient. The aerosol container is generally designed to deliver a predetermined dose of medicament upon each actuation by means of the valve, which can be opened either by depressing the valve while the container is held stationary or by depressing the container while the valve is held stationary.

Where the medicament container is an aerosol container, the valve typically comprises a valve body having an inlet port through which a medicament aerosol formulation may enter said valve body, an outlet port through which the aerosol may exit the valve body and an open/close mechanism by means of which flow through said outlet port is controllable.

The valve may be a slide valve wherein the open/close mechanism comprises a sealing ring and receivable by the sealing ring a valve stem having a dispensing passage, the valve stem being slidably movable within the ring from a valve-closed to a valve-open position in which the interior of the valve body is in communication with the exterior of the valve body via the dispensing passage.

Typically, the valve is a metering valve. The metering volumes are typically from 10 to 100 μl, such as 25 μl, 50 μl or 63 μl. Suitably, the valve body defines a metering chamber for metering an amount of medicament formulation and an open/close mechanism by means of which the flow through the inlet port to the metering chamber is controllable. Preferably, the valve body has a sampling chamber in communication with the metering chamber via a second inlet port, said inlet port being controllable by means of an open/close mechanism thereby regulating the flow of medicament formulation into the metering chamber.

The valve may also comprise a ‘free flow aerosol valve’ having a chamber and a valve stem extending into the chamber and movable relative to the chamber between dispensing and non-dispensing positions. The valve stem has a configuration and the chamber has an internal configuration such that a metered volume is defined there between and such that during movement between is non-dispensing and dispensing positions the valve stem sequentially: (i) allows free flow of aerosol formulation into the chamber, (ii) defines a closed metered volume for pressurized aerosol formulation between the external surface of the valve stem and internal surface of the chamber, and (iii) moves with the closed metered volume within the chamber without decreasing the volume of the closed metered volume until the metered volume communicates with an outlet passage thereby allowing dispensing of the metered volume of pressurized aerosol formulation. A valve of this type is described in U.S. Pat. No. 5,772,085. Additionally, intra-nasal delivery of the present compounds is effective.

To formulate an effective pharmaceutical nasal composition, the medicament must be delivered readily to all portions of the nasal cavities (the target tissues) where it performs its pharmacological function. Additionally, the medicament should remain in contact with the target tissues for relatively long periods of time. The longer the medicament remains in contact with the target tissues, the medicament must be capable of resisting those forces in the nasal passages that function to remove particles from the nose. Such forces, referred to as ‘mucociliary clearance’, are recognized as being extremely effective in removing particles from the nose in a rapid manner, for example, within 10-30 minutes from the time the particles enter the nose.

Other desired characteristics of a nasal composition are that it must not contain ingredients which cause the user discomfort, that it has satisfactory stability and shelf-life properties, and that it does not include constituents that are considered to be detrimental to the environment, for example ozone depletors.

A suitable dosing regime for the formulation of the present invention when administered to the nose would be for the patient to inhale deeply subsequent to the nasal cavity being cleared. During inhalation the formulation would be applied to one nostril while the other is manually compressed. This procedure would then be repeated for the other nostril.

In one embodiment, the means for applying a formulation of the present invention to the nasal passages is by use of a pre-compression pump. Most preferably, the pre-compression pump will be a VP7 model manufactured by Valois SA. Such a pump is beneficial as it will ensure that the formulation is not released until a sufficient force has been applied, otherwise smaller doses may be applied. Another advantage of the pre-compression pump is that atomisation of the spray is ensured as it will not release the formulation until the threshold pressure for effectively atomising the spray has been achieved. Typically, the VP7 model may be used with a bottle capable of holding 10-50 ml of a formulation. Each spray will typically deliver 50-100 μl of such a formulation, therefore, the VP7 model is capable of providing at least 100 metered doses.

Spray compositions for topical delivery to the lung by inhalation may for example be formulated as aqueous solutions or suspensions or as aerosols delivered from pressurised packs, such as a metered dose inhaler, with the use of a suitable liquefied propellant. Aerosol compositions suitable for inhalation can be either a suspension or a solution and generally contain the compound of Formula (I) optionally in combination with another therapeutically active ingredient and a suitable propellant such as a fluorocarbon or hydrogen-containing chlorofluorocarbon or mixtures thereof, particularly hydrofluoroalkanes, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetra-fluoroethane, especially 1,1,1,2-tetrafluoroethane, 1,1,1,2,3,3,3-heptafluoro-n-propane or a mixture thereof. Carbon dioxide or other suitable gas may also be used as propellant. The aerosol composition may be excipient free or may optionally contain additional formulation excipients well known in the art such as surfactants, e.g., oleic acid or lecithin and cosolvents, e.g. ethanol. Pressurized formulations will generally be retained in a canister (e.g. an aluminum canister) closed with a valve (e.g. a metering valve) and fitted into an actuator provided with a mouthpiece.

Medicaments for administration by inhalation desirably have a controlled particle size. The optimum particle size for inhalation into the bronchial system is usually 1-10 μm, preferably 2-5 μm. Particles having a size above 20 μm are generally too large when inhaled to reach the small airways. To achieve these particle sizes the particles of the active ingredient as produced may be size reduced by conventional means e.g., by micronization. The desired fraction may be separated out by air classification or sieving. Suitably, the particles will be crystalline in form. When an excipient such as lactose is employed, generally, the particle size of the excipient will be much greater than the inhaled medicament within the present invention. When the excipient is lactose it will typically be present as milled lactose, wherein not more than 85% of lactose particles will have a MMD of 60-90 μm and not less than 15% will have a MMD of less than 15 μm.

Intranasal sprays may be formulated with aqueous or non-aqueous vehicles with the addition of agents such as thickening agents, buffer salts or acid or alkali to adjust the pH, isotonicity adjusting agents or anti-oxidants.

Solutions for inhalation by nebulization may be formulated with an aqueous vehicle with the addition of agents such as acid or alkali, buffer salts, isotonicity adjusting agents or antimicrobials. They may be sterilised by filtration or heating in an autoclave, or presented as a non-sterile product.

For all methods of use disclosed herein for the compounds of Formula (I), the daily topical dosage regimen will preferably be from 0.01 mg to 1000 mg, administered one to four times daily. The daily inhalation dosage regimen will preferably be from about 0.05 microgram/kg to about 1 mg/kg per day, more preferably from about 0.2 microgram/kg to about 20 microgram/kg, administered in one or more daily doses. The daily intranasal dosage regimen will preferably be from about 0.05 microgram/kg to about 1 mg/kg per day, more preferably from about 0.2 microgram/kg to about 20 microgram/kg, administered in one or more daily doses. It will also be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of Formula (I) or a pharmaceutically acceptable salt thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill in the art that the optimal course of treatment, i.e., the number of doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof given per day for a defined number of days, can be ascertained by those skilled in the art using conventional course of treatment determination tests.

The novel compounds of Formula (I) may also be used in association with the veterinary treatment of mammals, other than humans, in need of antagonism of a muscarinic receptor or a PDE4 enzyme. In particular, the treatment, therapeutically or prophylactically, in animals include disease states such as those noted herein in the Methods of Treatment section.

For use herein treatment may include prophylaxis. It may also include reducing the symptoms of, ameliorating the symptoms of, reducing the severity of, reducing the incidence of, or any other change in the condition of the patient, which improves the therapeutic outcome.

It should be understood that in addition to the ingredients particularly mentioned above, the formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents, or those for inhalation may include carriers, such as lactose.

The anticipated therapeutic activity for a dual pharmacophore antagonist of muscarinic receptors and an inhibitor of the PDE4 enzyme within a single molecule is both as a bronchodilator (provided by both muscarinic receptor antagonist activity and PDE4 inhibition) and as an anti-inflammatory (by elevation of cytosolic levels of 3′,5′-cyclic adenosine monophosphate (cAMP) through inhibition of the PDE4 enzyme and by blockade other pro-inflammatory mechanisms mediated through muscarinic receptors on immune and resident cells) within the lungs. There is also a potential for further positive cooperatively as an anti-inflammatory through simultaneous interaction of downstream signaling pathways via modulation of both targets within the same cell.

Muscarinic receptors are coupled to G-proteins (M₁, M₃ & M₅ via G_(q/11) and M₂ & M₄ via G_(i/0)) which can lead to activation of a number of intracellular targets and signaling cascades. For example, M₂ and M₄ receptors via G_(i/0) can decrease cellular adenylyl cyclase levels and increase MAP kinase activation whereas M₁, M₃ & M₅ receptors via G_(q/11) can elevate phospholipase Cβ (PLCβ) and increase MAP kinase activation (Nathanson N M. A multiplicity of muscarinic mechanisms: enough signaling pathways to take your breathe away. Proc. Natl. Acad. Sci. USA. 2000; 97:6245-6247. Lanzafame AA. Cellular signaling mechanisms for muscarinic acetylcholine receptors. Recept. Chann. 2003; 9:241-260).

There is a potential therefore to elevate intracellular levels of cAMP through inhibition of the PDE4 enzyme, the enzyme responsible for breaking down cAMP into 5′-AMP and by increasing adenylyl cyclase activity, the enzyme responsible for conversion of ATP into cAMP, via blockade of M₂ receptors on immune cells, thus inhibiting acetylcholine signaling through G_(i/0) and therefore inhibiting decrease of adenylyl cyclase activity. Simultaneous activities of M₂ receptor blockade and PDE4 inhibition at the same cell would therefore lead to elevation of intracellular cAMP by two independent mechanisms increasing the overall concentration of cAMP within the cell. Elevated levels of cyclic AMP has been shown to have anti-inflammatory activity in a range of immune cells including T-cells, macrophages and neutrophils as well as resident lung cells such as epithelial and airway smooth muscle cells. Elevated cAMP can also cause airway smooth muscle relaxation and may offer a further mechanism independent of M₃ receptor blockade to initiate bronchodilation. For a full review of the potential therapeutic activities of PDE4 inhibitors in respiratory diseases see: Kroegel C & Foerster M. Phophodiesterase-4 inhibitors as a novel approach for the treatment of respiratory disease: cilomilast. Expert Opin. Investig. Drugs 2007; 16:109-124. Dastidar SG. et al., Therapeutic benefit of PDE4 inhibitors in inflammatory diseases. Curr. Opin. Investig. Drugs 2007; 8:364-372. Krymskaya VP & Panettieri RA. Phosphodiesterases regulate airway smooth muscle function in health and disease Curr. Top. Dev. Biol. 2007; 79:61-74. Spina D. The potential of PDE4 inhibitors in respiratory disease. Curr. Drug Targets Inflamm. Allergy 2004; 3:231-236.

The disposition within the lungs of a single drug substance which acts at both muscarinic receptors and as a PDE4 inhibitor at the same cell offers the greatest opportunity for cooperative anti-inflammatory or bronchodilator activity through modulation of these independent targets. This approach offers a greater potential to maximize the interaction of these two independent mechanisms compared to co-administration of two pharmacophores directed against each target as co-disposition at the cells of the lungs cannot be guaranteed through the second approach. The novel single dual pharmacophore approach outlined here, therefore offers a significantly greater potential for co-disposition to cells of the lung compared to administration of two separate pharmacophores directed against each target. Further to this such a pharmacophore will also be more amenable to combination with existing or other novel inhaled therapies for the treatment of respiratory diseases.

Therefore, compounds and pharmaceutical formulations according to the invention may be used in combination with or include one or more other therapeutic agents, for example selected from anti-inflammatory agents, other selective anticholinergic agents (particularly an M₁, M₂, or M₁/M₂ receptor antagonist), β₂-adrenoreceptor agonists, antiinfective agents (e.g. antibiotics, antivirals), or antihistamines. The invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof together with one or more other therapeutically active agents, for example selected from an anti-inflammatory agent (for example a corticosteroid or an NSAID), an anticholinergic agent, β₂-adrenoreceptor agonist, an antiinfective agent (e.g. an antibiotic or an antiviral), or an antihistamine. One aspect of the present invention are combinations comprising a compound of Formula (I) or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof together with a corticosteroid, and/or an anticholinergic, and/or a PDE4 inhibitor. Preferred combinations are those comprising one or two other therapeutic agents.

It will be clear to a person skilled in the art that, where appropriate, the other therapeutic ingredient(s) may be used in the form of salts, (e.g. as alkali metal or amine salts or as acid addition salts), or prodrugs, or as esters (e.g. lower alkyl esters), or as solvates (e.g. hydrates) to optimize the activity and/or stability and/or physical characteristics (e.g. solubility) of the therapeutic ingredient. It will be clear also that where appropriate, the therapeutic ingredients may be used in optically pure form.

One suitable combination of the present invention comprises of compound of the invention together with a β₂-adrenoreceptor agonist.

Examples of β₂-adrenoreceptor agonists include salmeterol (which may be a racemate or a single enantiomer, such as the R-enantiomer), salbutamol, formoterol, salmefamol, fenoterol or terbutaline and salts thereof, for example the xinafoate salt of salmeterol, the sulphate salt or free base of salbutamol or the fumarate salt of formoterol. Long-acting β₂-adrenoreceptor agonists are preferred, especially those having a therapeutic effect over a 24 hour period, such as salmeterol or formoterol.

Suitable long acting β₂-adrenoreceptor agonists include those described in WO02/66422A, WO02/270490, WO02/076933, WO03/024439, WO03/072539, WO 03/091204, WO04/016578, WO04/022547, WO04/037807, WO04/037773, WO04/037768, WO04/039762, WO04/039766, WO01/42193 and WO03/042160, whose disclosures are incorporated by reference herein.

Preferred long-acting β₂-adrenoreceptor agonists are:

-   3-(4-{[6-({(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]ethyl}amino)     hexyl]oxy}butyl)benzenesulfonamide; -   3-(3-{[7-({(2R)-2-hydroxy-2-[4-hydroxy-3-hydroxymethyl)phenyl]ethyl}-amino)heptyl]oxy}propyl)benzenesulfonamide; -   4-{(1R)-2-[(6-{2-[(2,6-dichlorobenzyl)oxy]ethoxy}hexyl)amino]-1-hydroxyethyl}-2-(hydroxymethyl)phenol; -   4-{(1R)-2-[(6-{4-[3-(cyclopentylsulfonyl)phenyl]butoxy}hexyl)amino]-1-hydroxyethyl}-2-(hydroxymethyl)phenol; -   N-[2-hydroxyl-5-[(1R)-1-hydroxy-2-[[2-4-[[(2R)-2-hydroxy-2-phenylethyl]amino]phenyl]ethyl]amino]ethyl]phenyl]foramide,     and -   N-2{2-[4-(3-phenyl-4-methoxyphenyl)aminophenyl]ethyl}-2-hydroxy-2-(8-hydroxy-2(1H)-quinolinon-5-yl)ethylamine.

Suitable anti-inflammatory agents include corticosteroids. Suitable corticosteroids which may be used in combination with the compounds of the invention are those oral and inhaled corticosteroids and their pro-drugs which have anti-inflammatory activity. Examples include methyl prednisolone, prednisolone, dexamethasone, fluticasone propionate, 6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioic acid S-fluoromethyl ester, 6α,9α-difluoro-11β-hydroxy-16α-methyl-3-oxo-17α-propionyloxy-androsta-1,4-diene-17β-carbothioic acid S-(2-oxo-tetrahydro-furan-3S-yl) ester, 6α,9α-difluoro-11β-hydroxy-16α-methyl-17α-(1-methylcylopropylcarbonyl)oxy-3-oxo-androsta-1,4-diene-17β-carbothioic acid S-fluoromethyl ester, 6α,9α-difluoro-11β-hydroxy-16α-methyl-3-oxo-17α-(2,2,3,3-tetramethylcyclopropylcarbonyl)oxy-androsta-1,4-diene-17β-carboxylic acid cyanomethyl ester, beclomethasone esters (such as the 17-propionate ester or the 17,21-dipropionate ester), budesonide, flunisolide, mometasone esters (such as the furoate ester), triamcinolone acetonide, rofleponide, ciclesonide, (16α,17-[[(R)-cyclohexylmethylene]bis(oxy)]-11β,21-dihydroxy-pregna-1,4-diene-3,20-dione), butixocort propionate, RPR-106541, and ST-126. Preferred corticosteroids include fluticasone propionate, 6α,9α-difluoro-11β-hydroxy-16α-methyl-17α-[(4-methyl-1,3-thiazole-5-carbonyl)oxy]-3-oxo-androsta-1,4-diene-17β-carbothioic acid S-fluoromethyl ester and 6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioic acid S-fluoromethyl ester, more preferably 6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioic acid S-fluoromethyl ester.

Non-steroidal compounds having glucocorticoid agonism that may possess selectivity for transrepression over transactivation and that may be useful in combination therapy include those covered in the following patents: WO03/082827, WO01/10143, WO98/54159, WO04/005229, WO04/009016, WO04/009017, WO04/018429, WO03/104195, WO03/082787, WO03/082280, WO03/059899, WO03/101932, WO02/02565, WO01/16128, WO00/66590, WO03/086294, WO04/026248, WO03/061651, WO03/08277.

Suitable NSAID's include sodium cromoglycate, nedocromil sodium, leukotriene antagonists, inhibitors of leukotriene synthesis (for example, montelukast), iNOS inhibitors, tryptase and elastase inhibitors, beta-2 integrin antagonists and adenosine receptor agonists or antagonists (for example, adenosine 2a agonists), cytokine antagonists (for example, chemokine antagonists, such as a CCR3 antagonist) or inhibitors of cytokine synthesis, 5-lipoxygenase inhibitors, p38 inhibitors, and IKK2 inhibitors. Suitable other β₂-adrenoreceptor agonists include salmeterol (for example, as the xinafoate), salbutamol (for example, as the sulphate or the free base), formoterol (for example, as the fumarate), fenoterol or terbutaline and salts thereof. An iNOS (inducible nitric oxide synthase inhibitor) is preferably for oral administration. Suitable iNOS inhibitors include those disclosed in WO93/13055, WO98/30537, WO02/50021, WO95/34534 and WO99/62875. Suitable CCR3 inhibitors include those disclosed in WO02/26722.

Suitable antihistamines (also referred to as H₁-receptor antagonists) include any one or more of the numerous antagonists known which inhibit H₁-receptors, and are safe for human use. All are reversible, competitive inhibitors of the interaction of histamine with H₁-receptors. The majority of these inhibitors, mostly first generation antagonists, are generally represented by three types of antihistamines: ethanolamines, ethylenediamines, and alkylamines. In addition, other first generation antihistamines include those which can be characterized as based on piperizine and phenothiazines. Second generation antagonists, which are non-sedating, have a similar structure-activity relationship in that they retain the core ethylene group (the alkylamines) or mimic the tertiary amine group with piperizine or piperidine. Exemplary antagonists are as follows:

Ethanolamines: carbinoxamine maleate, clemastine fumarate, diphenylhydramine hydrochloride, and dimenhydrinate.

Ethylenediamines: pyrilamine maleate, tripelennamine HCl, and tripelennamine citrate.

Alkylamines: chloropheniramine and its salts such as the maleate salt, and acrivastine.

piperazines: hydroxyzine HCl, hydroxyzine pamoate, cyclizine HCl, cyclizine lactate, meclizine HCl, and cetirizine HCl.

Piperidines: Astemizole, levocabastine HCl, loratadine or its descarboethoxy analogue, and terfenadine and fexofenadine hydrochloride or another pharmaceutically acceptable salt.

The combinations referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above together with a physiologically acceptable diluent or carrier represent a further aspect of the invention.

The individual compounds of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations. Appropriate doses of known therapeutic agents will be readily appreciated by those skilled in the art.

The invention will now be described by reference to the following biological examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention.

BIOLOGICAL EXAMPLES

As compounds of Formula (I) have dual pharmacophores, maximizing both activities is a component of the testing process. There is a desire to balance the antagonism of the muscarinic M₃ receptor with the inhibition of the PDE4 enzyme. Although the M₃ antagonism is measured in a human receptor expressed in a mammalian cell line as described herein, PDE4 is usually measured on isolated human enzyme, therefore a secondary cell assay is monitored that reflects intracellular PDE4 inhibition. An example of such a cellular assay is the PBMC assay as shown below. Therefore it is desired to optimize PDE4 inhibition in the cell (measured using PBMC assay). Desired attributes of the molecule would be to maintain or improve the M₃ pharmacophores potency with no or partial M₁ agonism since agonism of the M₁ receptor is usually counter-indicated. Another attribute is to decrease the dropoff between the PDE4 enzyme assay and inhibition reflected in the PBMC assay. Since both pharmacophores are in a single molecule, it is desirable to enhance intracellular inhibition of PDE4 reflected in the PBMC assay while retaining significant activity against the transmembrane M₃ receptor. In addition, in vivo efficacy and duration of action is not always reflected by in vitro measurements of activity, therefore other physiochemical properties of the molecules may be important for balanced efficacy at both targets. Therefore, one embodiment of the invention are compounds which posses appropriately balanced pharmacology, and have desirable physicochemical properties, such as solubility, dissolution rate, permeability, crystallinity, micronizability, and excipient compatibility. If the compounds are administered by inhalation, then low aqueous solubility is generally not suitable for a nebulized/solution formulation.

One embodiment of the invention is a display of sufficient antagonism at the M₃ receptor wherein p IC₅₀≧8.0 and a pA₂≧8.0, as well as inhibition of the PDE4 enzyme with a pIC₅₀≧8.0 and cellular activity (as reflected in the PBMC assay) with a pIC₅₀≧7.0.

In one embodiment of the invention compounds of Formula (I) are generally selective against agonism or partial agonism of the various muscarinic receptors (M₁, M₂, M₃) and PDE4>100-fold vs. other PDEs.

The inhibitory effects of compounds at the mAChR (muscarinic) receptor and the PDE4 enzyme for the present invention are determined by the following in vitro and in vivo functional assays.

mAChR (Muscarinic)Receptor Assays

In Vitro Assays

Muscarinic Receptor Radioligand Binding Assays

Radioligand Binding Studies to Determine Interaction at Cloned Human Receptors

The human M₁-M₃ receptors are cloned and stably expressed in Chinese Hamster Ovary (CHO) cell lines. M₂ ACh receptor is co-expressed with the chimeric G protein, Gqi5, in CHO cells. Competition for [³H]-N-methyl scopolamine (0.5 nM) binding is performed using crude CHO cell membranes using a Scintillation Proximity Assay (SPA). Atropine is run in every assay as the control.

In the SPA assay membranes are preincubated with wheatgerm agglutinin beads (GE) in 50 mM HEPES buffer (Sigma, St. Louis Mo.) (pH 7.4) at 4° C. for 30 min, and then incubated with 0.5 nM [³H]-N-methyl scopolamine (PerkinElmer) in a 96-well Optiplate (Perkin Elmer), for 2 hr in the presence of vehicle (1% DMSO) or compound (0.01-1000 nM), in 0.2 mL final volume, at room temperature. At the end of the incubation the plates are centrifuged (Beckman CS-6R) for 5 min at 2000 RPM, and counted in a Top Count Microplate Scintillation counter (model A9912 Packard, Meriden Conn.).

Concentration-response curves for each compound are run using duplicate samples in 3 independent experiments. Specific binding is determined by subtracting non-specific binding (defined in the presence of 0.3 μM Atropine) from total binding. IC₅₀ values are estimated from concentration-response curves and used to determine the inhibition constant (Ki) of each inhibitor using the Cheng and Prusoff equation [for competitive antagonists: The Kd's utilized for the calculations are: 0.17, 0.28, and 0.16, nM for M1, M2 and M3 respectively.

$K_{i} = \left. \rightarrow\frac{{IC}_{50}}{{\lbrack L\rbrack\text{/}K_{d}} + 1} \right.$ Membrane Preparation

Cells are harvested by centrifugation at 1000×g for 10 min at 4° C. The cell pellet is washed with Phosphate Buffered Saline (PBS) and quick frozen with liquid nitrogen. The pellet is stored at −80° C. until the membrane preparation is made. The frozen pellet is thawed and re-suspended in cold hypotonic membrane buffer (40 mM Tris, pH 7.5, 1 mM MgSO₄, 0.5 mM EDTA, 1 mM phenylmethylsulfonyl fluoride, 2.5 mg/L leupeptin, 0.1 mg/mL aprotinin) and incubated on ice for 5 min. The cell suspension is homogenized in a 40 mL Dounce homogenizer and centrifuged at 2000 rpm at 4° C. for 6 min to remove nuclei and cellular debris. The 2000 rpm pellet is resuspended in homogenization buffer and spun again at 2000 rpm for 6 min. This process is repeated two more times. The combined supernatant is collected and cell membranes are pelleted at 100000×g for 1 hr at 4° C. The membrane pellet is resuspended in membrane buffer and aliquots stored at −80° C. Protein concentration is quantified using the Bio-Rad protein assay reagent.

Calcium Mobilization Studies (FLIPR)

Studies to determine the effectiveness of antagonists to cause blockade of functional intracellular calcium fluxes following agonist (ACh) treatment of cloned human receptors.

This system is used for characterizations of antagonist-receptor interactions using four distinct variations of the FLIPR methodology: (a) potency: IC₅₀ determination, (b) potency: pA₂ determination, (c) reversibility of antagonist-receptor interaction, or (d) confirmation of no functional agonist activity.

-   Cell Source The human M1-M3 receptors are cloned and stably     expressed in Chinese Hamster Ovary (CHO) cells. The M2 receptors are     co-expressed with the chimeric G protein, Gqi5. -   Cell lines: M1 stable: Biocat #1044; M2+Gqi5 stable: Biocat #95663;     M3 stable: Biocat #1049 -   Method of Culture: CHO-M1, CHO-Gqi5-M2 and CHO-M3 cells are cultured     to confluence at 37° C. in a humidified incubator with 5% CO₂/95%     air. CHO-M1 and CHO-M3 are cultured in Alpha MEM with nucleosides     and L-glutamine and 10% fetal calf serum. Cells expressing the M2     receptor are cultured in DMEM/F12 media, supplemented with 200 mg/L     G418 (geneticin), and 10% fetal calf serum. -   Assay Readout: Calcium mobilization, monitored as change in     cytosolic calcium concentration, is measured as change in 516 nm     emission fluorescence intensity of cytosolic loaded Fluo-4, a green     fluorescent calcium indicator which exhibits large (>100-fold)     fluorescence intensity increases on binding to calcium, the change     in intensity being, therefore, directly related to cytosolic calcium     levels. The emitted fluorescence from all 96 wells is measured     simultaneously using a cooled CCD camera. Data points are collected     every second. Maximal change in emission from each well after     simultaneous addition of agonist or compound to each of the 96 wells     is then exported to an excel spreadsheet. This data is then     transferred to GraphPad Prism Version 4.03 for plotting of response     to each treatment condition (ACh or compound).     Experimental Protocols: -   Cell plating: A microtiter plate based calcium mobilization FLIPR     (Fluorometric Imaging Plate Reader, Molecular Devices, Sunnyvale     Calif., [Schroeder KS, Neagle, BD. FLIPR: a new instrument for     accurate, high throughput optical screening. J. Biomol. Screen.     1996; 1:75.]) assay is used for the functional characterization of     compounds against M1, M2 (w/Gqi5) and M3 ACh receptors stably     expressed in CHO cells. On the day prior to assay, cells are plated     in 96 well, blackwall, clear bottom plates (Packard View) at a     concentration of 40000 cells per well and incubated at 37° C. in a     humidified incubator with 5% CO₂/95% air for 18 to 24 hours.     a) IC₅₀ Determination for Antagonists: -   Receptor antagonist characterization (IC₅₀ determination), compounds     tested for potency of inhibition of ACh induced muscarinic receptor     activation: To evaluate antagonist potency of compounds against the     M₁, M₂ and M₃ receptors, cell culture media is aspirated and     replaced with 100 μL of dye load media [Eagles Minimal Essential     Medium (EMEM) with Earl's salts and L-Glutamine, 0.1% BSA     (Seriologicals Corporation), 4 μM Fluo-4-acetoxymethyl ester     fluorescent indicator dye (Fluo-4 AM, Molecular Probes, Eugene,     Oreg.) and 2.5 mM probenecid]. Cells are then incubated for 1 hour     at 37° C. The dye load media is then aspirated off the cells and     replaced with identical media without Fluo-4 AM and with 0.1%     Gelatin (BSA removed) and 2.5 mM probenecid. Cells are incubated for     10 minutes at 37° C. and then washed 3 times with KRH assay buffer     [Krebs Ringer Henseleit (120 mM NaCl, 4.6 mM KCl, 1.03 mM KH₂PO₄, 25     mM NaHCO₃, 1.0 mM CaCl₂, 1.1 mM MgCl₂, 11 mM Glucose, 20 mM HEPES     (pH 7.4)) with 0.1% gelatin and 2.5 mM probenecid]. 100 μL KRH assay     buffer with 0.1% gelatin and 2.5 mM probenecid is added to wells of     dye loaded and washed cells followed by 50 μL of 3×compound     (1×10⁻⁸-3.3×10⁻⁵ M final in the assay) and plate warmed to 37° C.     for 10 minutes before being placed in FLIPR where the dye loaded,     compound pretreated cells are exposed to excitation light (488 nm)     from a 6 watt Argon Laser. The basal emission fluorescence is     measured, then the cellular response to an EC₈₀ concentration of ACh     (3.3 nM against M₁, 10 nM against M₂ and 1.0 nM against M₃) prepared     in KRH assay buffer with 0.1% BSA (no gelatin), is monitored in     FLIPR for 90 seconds and then 50 μL of 100 μM ATP (assay     concentration of 20 μM) is added to check cell viability (H. M.     Sarau et al, 1999. Mol. Pharmacol. 56, 657-663). Maximal change in     emission from each well, vehicle or compound pretreated, after     simultaneous addition of ACh to each of the 96 wells is then     determined. The IC₅₀ is defined as the compound pretreatment     concentration which inhibits 50% of the ACh induced response.     Compounds are believed to be active in this assay are those having     an IC₅₀ of about 33 uM to about 10 nM or less. The IC₅₀ is defined     as the compound pretreatment concentration which inhibits 50% of the     ACh induced response.

A compound is believed to be active in this assay if it has an IC₅₀ of between 33 uM and 10 nM or less. Exemplary compounds of Formula (I) which have been tested in this assay and found to be the most active can be found in Examples 191-200, 202-226, 228, 230-234, 236-240, 242-269, 271, 273-280, 282-288, 290-293, 295-297, 300-301, 303-305, 307, 311, 314-315, 317-336, 338-340, 342-354, 356-361, and 363-433.

b) pA₂ Determination for Antagonists:

-   Single concentration kinetic characterization of compounds tested     for potency of inhibition of ACh induced muscarinic receptor     activation: pA₂: Compounds which show IC₅₀'s of <1.0 μM may be     further characterized in a single compound concentration kinetic     assay. To confirm antagonist potency of more potent compounds     against the M1, M2 and M3 receptors, dye loaded (culture media is     aspirated, replaced with 100 μL of dye load media and incubated for     1 hour at 37° C.) and washed cells (washed three times with 100 μL     KRH assay buffer) are treated with 150 μL of KRH assay buffer with     0.1% gelatin and 2.5 mM probenecid containing vehicle (0.01% DMSO),     for control response, or appropriate concentration of antagonist     (single concentration for each column of 12 wells, concentration     determined from IC₅₀ value) and incubated for 20 minutes at 37° C.     Buffer is aspirated off and 150 μL of fresh KRH assay buffer with     0.1% gelatin and 2.5 mM probenecid containing vehicle (0.01% DMSO)     or appropriate concentration of compound is added and incubated for     10 minutes at 37° C. Plates are then placed into FLIPR for     fluorescent measurements. After determination of basal fluorescence     emission, a concentration range of ACh (0.033-100,000 nM for M1/M3     and 0.33-1,000,000 nM for M2) is added to vehicle or compound     treated (columns of 12 wells) cells to determine the shift of     receptor potency in response to ACh in presence of compound.     Compound potency at the receptor is determined using the following     formula: pA₂=log(DR-1)−log[B] where DR is the dose ratio defined as     the ratio of equiactive concentration (EC₅₀) of agonist in presence     and absence of antagonist and [B] is concentration of antagonist.     c) Determination of Antagonist Reversibility -   Evaluation of antagonist-receptor occupancy following antagonist     wash-out (reversibility) using FLIPR methodology: After aspirating     off growth media the cells are washed 3 times with 100 μl KRH assay     buffer containing 0.1% gelatin. Each column (12 wells) is treated     with 150 μL of EMEM containing 0.1% gelatin with vehicle (0.01%     DMSO) or antagonist at an appropriate concentration: 1.0 nM, 10 nM,     100 nM or 1000 nM, (washout columns), or not treated (no washout     columns) and incubated for 60 minutes at 37° C. EMEM is aspirated     and KRH assay buffer containing 0.1% gelatin with vehicle (0.01%     DMSO) or antagonist is added to washout columns and incubated at     37° C. for 20 minutes. Buffer with vehicle or compound is aspirated     and cells retreated and incubated at 37° C. for an additional 10     minutes. Buffer with vehicle or compound is then aspirated and cells     washed 3 times with KRH assay buffer containing 0.1% BSA. KRH buffer     (100 μL) containing 0.1% BSA is then added and cells incubated for     30 minutes at 37° C. and washed 3 times. Cells are incubated for a     further 30 minutes and washed 3 times, followed by a further 30     minute incubation. After this 90 minute washout, all cells are     washed 3 times with KRH containing 0.1% gelatin. Cells are loaded     with dye using 150 μL dye load media with 0.1% gelatin and 2.5 mM     probenecid for washout columns or same dye load media with 0.1%     gelatin and 2.5 mM probenecid with vehicle (0.01% DMSO) or     appropriate concentration of compound (1.0 nM, 10 nM, 100 nM or 1000     nM) for no washout columns and incubated for 60 minutes at 37° C.     Dye load media is aspirated and cells are retreated with 150 μL of     KRH assay buffer containing 0.1% gelatin and 2.5 mM probenicid for     washout columns or KRH assay buffer containing 0.1% gelatin and 2.5     mM probenicid and vehicle or appropriate concentration of compound     for no washout columns. Cells are incubated for 20 minutes at 37° C.     Pretreatment buffer is aspirated and 150 μL of fresh KRH assay     buffer with 0.1% gelatin and 2.5 mM probenecid is then added to     washout columns and the same buffer containing vehicle (0.01% DMSO)     or the appropriate concentration of antagonist is added to the no     washout columns. Plates are incubated for 10 minutes at 37° C. and     plates placed into FLIPR where fluorescence is monitored. Baseline     measurements are recorded and acetylcholine concentration response     curves are added to each column while continuing to monitor     fluorescence. Comparison of ACh concentration response curves is     performed between vehicle-treated and antagonist-treated [1.0 nM, 10     nM, 100 nM or 1000 nM] cells following washout to determine if there     remained a shift in the EC₅₀ value post washout. Fold-shift (fs)     values were determined using the following formula: fs=[X]/[V];     wherein X is the concentration of acetylcholine required to elicit a     50% maximum calcium mobilization response following antagonist     treatment and washout; V is the concentration of acetylcholine     required to elicit a 50% maximum calcium mobilization response     following vehicle treatment and washout.     d) Confirmation of No Agonist Activity -   Receptor agonist characterization (EC₅₀ determination): compounds     tested to confirm no agonist potential at muscarinic receptors: To     evaluate agonist potential of compounds and ACh potency for the M₁,     M₂ and M₃ receptors, culture media is aspirated and replaced with     100 μL of dye load media. Cells are then incubated for 1 hour at     37° C. The dye load media is then aspirated off the cells and     replaced with identical media without Fluo-4 AM and with 0.1%     Gelatin (BSA removed) and 2.5 mM probenecid. Cells are incubated for     10 minutes at 37° C. and then washed 3 times with 100 μL KRH assay     buffer. 100 μL KRH assay buffer with 0.1% gelatin and 2.5 mM     probenecid is added to wells of dye loaded and washed cells and     plate warmed to 37° C. for 10 minutes before being placed in FLIPR     where dye loaded cells are exposed to excitation light (488 nm) from     a 6 watt Argon Laser. The basal emission fluorescence is measured,     then the cellular response to a concentration range of ACh or     compound (50 μL of 3× in assay buffer) is monitored in FLIPR for 90     seconds and then 50 μL of 100 μM ATP (assay concentration of 25 μM)     was added to check cell viability. The EC₅₀ is the ACh or compound     concentration required to obtain 50% the maximal response.

Superfusion Protocols

All procedures were performed in accredited facilities in accordance with Universal Precautions for Handling Human Blood, Body Fluids, and Tissue (BAR # 88-06-22-060) and institutional guidelines including the Guide for the Care and Use of Laboratory Animals (DHSS #NIH 85-23) and approved protocol #86-077 (Animal Care and Use Committee, GlaxoSmithKline). Human lungs from organ donors were obtained from the National Disease Research Interchange (NDRI, Philadelphia, Pa., www.ndriresource.org). Sections of bronchus were removed from the lung and cleaned of adherent connective, parenchymal and fatty tissue. Bronchial strips of approximately 3-4 mm in width were prepared and placed into modified Krebs-Henseleit solution. Composition of the solution was (mM): NaCl (113.0), KCl (4.8), CaCl₂ (2.5), KH₂PO₄ (1.2), MgSO₄ (1.2), NaHCO₃ (25.0) and dextrose (11.0) and equilibrated with 95% O₂: 5% CO₂ and maintained at 37° C.; meclofenamic acid (1 μM) was added to block endogenous cycloxygenase activity. Alternatively, trachea was removed from male Hartely guinea pigs (Charles River, Portage, Mich.; weight range 450-650 g). The epithelium of the trachea was removed and strips were cut, approximately 2 cartilage rings in width. Individual tissues were suspended via silk suture in a superfusion chamber (Coleman, 1989; Harvard Apparatus, Inc., Holliston, Mass., www.harvardapparatus.com) and connected to BIOPAC TSD125C transducers. The tissues were then continuously superfused with Krebs-Henseleit solution at 2 mL/min for the duration of the experiment. Stock solutions of agonist and antagonist were infused (0.02 mL/min) via 22-gauge needle inserted into the superfusion tubing. Mechanical responses were recorded isometrically using a commercially-available data acquisition system (MP100WS/Acknowledge; BIOPAC Systems, Goleta, Calif., www.biopac.com) interfaced with a computer.

Duration of PDE4M compound-induced inhibition of the carbachol response was investigated in two ways. The first protocol described was used to assess onset and offset of compound-induced inhibition. The second protocol was used to evaluate inhibition of the carbachol response in the presence of infused compounds as compared to inhibitory activity remaining after overnight washout.

Protocol A: Tissues were suspended under an optimal resting tension of 1.5 g. After a 60 min equilibration period, the tissues were contracted with carbachol (1 μM) for the duration of the experiment. Upon reaching a sustained contraction isoproterenol (10 μM) was administered to maximally relax the tissue, and this change served as a reference. Isoproterenol exposure was halted and the carbachol-induced tension allowed to recover. Compounds and vehicle were infused at a single concentration per tissue until a sustained level of inhibition was attained. Compounds were infused for six hours and upon which the infusion of compounds and vehicle was halted. Carbachol-induced tension in tissues was then allowed to recover for 10 hours. After this recovery period, carbachol was removed from the perfusate and tissues allowed to return to baseline tone. A carbachol concentration-response curve was then generated, whole-log increments from 10 nM to 100 μM, followed by a 1 mM histamine-induced contraction for reference.

The following parameters were determined for each concentration of antagonist, and expressed as the mean±SEM for n individual tissues (n=numbers). Inhibition of the carbachol-induced contraction was expressed as a percent of the isoproterenol reference response. The onset halftime to maximal inhibition of tension (ON t_(1/2)) was determined. The offset halftime of tension recovery (OFF t_(1/2)), following removal of the compound from the superfusate, was determined by measuring the time required for tension to return to the level used to measure the respective onset halftime. Tension recovery was plotted vs. time as a percentage of the % recovery of maximal inhibition. Post-recovery concentration-response curves were plotted with data as a percent of the 1 mM post-histamine reference contractions. EC₅₀ and fold-shift vs. control values were calculated for each compound tested.

Protocol B: Tissues were suspended under an optimal resting tension of 1.5 g. After an incubation period to reach stable basal tone, histamine (10 μM) was infused to assess tissue contraction response. After tension reached a plateau, histamine infusion was halted and tissues tension allowed to return to baseline. Compounds and vehicle were then infused onto the tissues for 6 hours. A carbachol concentration-response curve was generated, in the presence of infused compounds or vehicle, by infusing carbachol over the tissues in cumulative half-log increments, 10 nM to 100 μM, followed by a 1 μM histamine-induced contraction for reference. Upon completion of this curve, infusion of compounds into the perfusate was halted and tissue tension allowed to return to baseline. The tissues were then washed with perfusate buffer overnight. The following morning, histamine (10 μM) was again infused to contract the tissues and assess tissue response. After tension reached a plateau, histamine infusion was halted and tissues tension allowed to return to baseline. Another carbachol concentration-response curve was generated, this time in the absence of infused compounds other than that remaining after the overnight washout.

Agonist-induced responses for each tissue were expressed as a percentage of the reference histamine (10 uM)-induced contraction obtained at the end of the curve. Geometric mean EC₅₀ values were calculated from nonlinear regression analyses of data (Motulsky, 2003). EC₅₀ and fold-shift vs. control values were calculated for each compound tested. For tissues where carbachol concentration-response curves were generated in the presence of infused test compounds, antagonist potencies were calculated and expressed as pK_(B) and pA₂ where appropriate (Arunlakshana & Schild, 1958): pKB=−log [antagonist]/X−1, where X is the ratio of agonist concentration required to elicit 50% of the maximal contraction in the presence of the antagonist compared with that in its absence and pA₂=−log of the antagonist dissociation constant.

In Vivo Assays:

Inhibition of Acetylcholine-Induced Bronchoconstriction in Conscious Guinea Pigs

a. Method

Procedure for Wet Suspension Intratracheal Dosing.

A stock solution of 5% weight/volume of Tween 80 is made at least one day prior to dosing. The solution is made by dissolving 1 gram of Tween 80 in a total volume of 20 ml sterile saline. On the day of dosing the stock 5% Tween solution is diluted 1:10 in sterile saline for a final concentration of 0.5% Tween. This solution is filtered through a 0.22 micron syringe filter to yield the final wet vehicle. Animals are weighed and the weights averaged for dose calculations: ((animal weight [kg])×(dose in [mg/kg]))/(dose volume [ml])=Dose Concentration [mg/ml]

Drug is weighed and placed into a glass homogenizer with the appropriate amount of vehicle, i.e. if 1.5 mg is weighed, than 1 ml vehicle is be added. The mixture is then homogenized by hand until it appears uniform. For doses lower than 1.0 mg/kg appropriate dilution of the suspension is made immediately after homogenization.

A one ml syringe capped with a 22 ga 2.5 inch rat gavage needle is filled with 200 μl of dosing solution. After an animal is anesthetized with isoflorane they are placed in the supine position and the dosing needle is introduced into the trachea via the mouth. After the drug solution is injected into the trachea the animal is returned to a recovery cage. Recovery from anesthesia is noted within 5 minutes.

Whole Body Plethysmograph Determination of Penh in Conscious Guinea Pigs:

Four and 24 hours (for dose-response experiment) and 4, 24, 48 and 72 hours (for duration of action experiment) after intratracheal drug or vehicle administration, male Dunkin-Hartley guinea pigs (650-750 g) (Charles River Labs, St Constance Canada) are placed into a whole body plethysmograph box (internal volume of approximately 7 liters). A bias air flow of 2 L/minute was applied to the box and flow changes in the box are measured and recorded using a Buxco XA data acquisition and respiratory analysis system (Buxco Electronics, Wilmington, N.C.). Animals are allowed to acclimate to the plethysmograph box for 3 minutes before air flow data is recorded. Recordings ire collected for 5 minutes to determine basal airway parameters. Animals ire exposed to an aerosol of acetylcholine (ACh) produced by an ultrasonic nebulizer (Delvibiss Pulmosonic 5000D) (3.5 mg/mL, pushed by a trickle flow of 0.6 mL/minute for 36 seconds followed by a 2 minute drying time) that generates an aerosol into a mixing chamber, then directly into the plethysmographic box airstream. Measurements are collected for 10 minutes following the ACh exposure. Collected values are retained and Penh (enhanced pause) is calculated. Penh has previously been shown as an index of airway obstruction and correlates with increased intrapleural pressure (Hamelmann E. et al., Noninvasive measurement of airway responsiveness in allergic mice using barometric plethysmography. Am. J. Crit. Care Med. 156:766-75]. The algorithm for the Penh calculation is as follows: Penh=[(expiratory time/relaxation time)−1]×(peak expiratory flow/peak inspiratory flow) where relaxation time is the amount of time required for 70% of the tidal volume to be expired. Animals are returned to caging until the next noted exposure timepoint. Each animal's baseline airway parameter is used as its own control when determining the effect of ACh aerosol exposure.

PDE4 Assays

In Vitro Assays

Inhibition of Phosphodiesterase IVB Enzyme Activity

Human recombinant PDE4B, in particular the 2B splice variant thereof (HSPDE4B2B), is disclosed in WO 94/20079 and also in M. M. McLaughlin et al., “A low Km, rolipram-sensitive, cAMP-specific phosphodiesterase from human brain: cloning and expression of cDNA, biochemical characterization of recombinant protein, and tissue distribution of mRNA”, J. Biol. Chem., 1993, 268, 6470-6476. For example, in Example 1 of WO 94/20079, human recombinant PDE4B is described as being expressed in the PDE-deficient yeast Saccharomyces cerevisiae strain GL62. PDE4B expression is induced by the addition of 150 μM CuSO₄.

For luminescence-coupled assay based screening the supernatant fractions of yeast cell lysates are subjected to Cibacron blue affinity chromatography, dialysis and desalting, to enrich for PDE4B and to remove components, e.g. ATP, able to interfere with the assay. Human recombinant PDE4D (HSPDE4D3A) is disclosed in P. A. Baecker et al., “Isolation of a cDNA encoding a human rolipram-sensitive cyclic AMP phosphodiesterase (PDE IV_(D))”, Gene, 1994, 138, 253-256. Expression of human PDE4D in yeast, and subsequent preparation of the recombinant protein for assay was as described for PDE4B.

Inhibition of PDE Activity: Luciferase-Coupled PDE Assay

Inhibition of PDE4B and PDE4D are measured using a luminescence-coupled assay system developed by Cambrex. This assay system couples the formation of AMP, derived from PDE4-catalyzed hydrolysis of cAMP, to the formation of ATP. The ATP is then used as a substrate for Luciferase and results in light as a signal output. When PDE is inhibited or inactive, no AMP is produced, the Luciferase is inactive, and no light signal is produced. This assay is used in a quenched assay format, where PDE4 enzyme (2.5 μL; ˜120 μM enzyme in 40 mM Tris-HCl, 10 mM MgCl₂, 1 mM CHAPS, 0.01% BSA, pH 7.5.) and cAMP substrate (2.51 μL; 2 μM cAMP in 40 mM Tris-HCl, 1 mM MgCl₂, 1 mM CHAPS, 0.01% BSA, pH 7.5.) are added sequentially to a 384 well assay plate (Greiner 784075) pre-stamped with 12.5-50 mL compound at the desired concentration. The reaction is incubated at room temperature for 1 hr, then is quenched by the addition of enzyme stop solution (1.5 μL; prepared as described by vendor; catalog #LT27-253) and then the light signal is generated by the addition of detection reagent (2.5 μL, prepared as described by vendor, catalog # LT27-250). The luminescence is then measured on a Viewlux imager (Perkin Elmer) using emission filters of 613/55 nm or 618/40 nm and a 5 second exposure. Compounds are prepared in neat DMSO at a concentration of 10 mM. For inhibition curves, compounds are diluted using a three fold serial dilution and tested at 11 concentrations (e.g. 50 μM-0.8 nM or 25 μM-0.42 nM or 2.5 μM to 42 pM). Curves were analyzed using ActivityBase and XL fit, and results are expressed as pIC₅₀ values.

Compounds having a pIC₅₀ of about 5 or greater are believed to be active in this assay, with the upper limit of resolution being approximately pIC₅₀=10.2.

Curves were analyzed using ActivityBase and XLfit, and results are expressed as pIC₅₀ values. A compound was believed to be active in this assay if it had a pIC₅₀ of about 6 to 10.4 or greater against PDE4B. Compounds of Formula (I) which have been tested in the PDE4B assay and found to be the most active can be found in Examples 191-257, 259-311, 313-351, and 353-433.

Other In Vitro Assays:

Inhibition of TNF-α (TNF-Alpha) Production in Human PBMC (peripheral Blood Mononuclear Cell) Assay

A 96-well flat bottom polystyrene tissue culture plate (manufacturer code 167008 Thermo Fisher Scientific, Kamstrupvej 90, Kamstrup, Roskilde DK-4000 Denmark) is prepared by initially adding to column 1 ca. 10 mM of test compound dissolved in DMSO, which is diluted about 7.94 fold in the well with DMSO to give a 1.26 mM solution. For a more potent compound, a more diluted solution in DMSO may be used. The compound is further diluted with DMSO into columns 2 to 9 by 8 successive 3-fold dilutions using the Biomek® 2000 Laboratory Automation Workstation (Beckman Coulter, Inc., 4300 N. Harbor Boulevard, P.O. Box 3100, Fullerton, Calif. 92834-3100 USA). Column 10 is used as a DMSO negative control (High Signal, 0% response), while column 11, which contains 1.26 mM of the PDE4 inhibitor roflumilast, is used as a positive control (Low Signal, 100% response). About 1 μl (about 1 ul) of compound is transferred to the compound plate using a Biomek® FX Laboratory Automation Workstation.

PBMC cells (peripheral blood mononuclear cells) are prepared from heparinised human blood (using 1% v/v Heparin Sodium 1000 IU/ml Endotoxin Free, Leo Laboratories Ltd., Cashel Road, Dublin 12. Ireland, Cat No: PL0043/0149) from normal volunteers using the Accuspin™ System-Histopaque®-1077 essentially (Sigma-Aldrich Company Ltd., The Old Brickyard New Rd, Gillingham Dorset SP8 4XT). About 20 ml of blood is overlaid onto 15 ml Histopaque® in Accuspin™ tubes. The tube is then centrifuged at about 800 g for ca. 20 minutes. The cells are collected from the cell layer, washed by centrifugation (ca. 1300 g, ca. 10 minutes) and resuspended in RPMI1640 medium (Low endotoxin RPMI1640 medium, Cat No: 31870, Invitrogen Corporation Invitrogen Ltd, 3 Fountain Drive, Inchinnan Business Park, Paisley Pa. 4 9RF, UK) containing 10% foetal calf serum, 1% L-glutamine (Invitrogen Corporation, Cat No: 25030) and 1% penicillin/streptomycin (Invitrogen Corporation, Cat No: 15140). Viable cells are counted by trypan blue staining and diluted to 1×10⁶ viable cells/ml. About 50 μl (about 50 ul) of diluted cells and about 75 μl (about 75 ul) of LPS (ca. 1 ng/ml final; Sigma Cat No: L-6386) are added to the compound plate, which is then incubated at 37° C., 5% CO₂, for about 20 hours.

The supernatant is removed and the concentrations of TNF-α are determined by electrochemiluminescence assay using the Meso Scale Discovery (MSD) technology (Meso Scale Discovery, 9238 Gaither Road, Gaithersburg, Md. 20877, USA). See the “TNF-α (TNF-alpha) MSD Assay” described below for typical details.

Results can be expressed as pIC50 values for inhibition of TNF-α (TNF-alpha) production in PBMCs, and it should be appreciated that these results can be subject to variability or error.

TNF-α (TNF-Alpha) MSD Assay:

-   MSD Human Serum Cytokine Assay Diluent, (25 μl) Meso Scale     Discovery, 9238 Gaither Road, Gaithersburg, Md. 20877) is added to a     96-well High-Bind MSD plate pre-coated with anti-hTNF alpha capture     antibody (MA6000) and then incubated for about 24 hours at 4° C. to     prevent non-specific binding. About 20 μl (ul) of supernatant from     the PBMC plate are then transferred from columns 1-11 to columns     1-11 of the MSD plate using the Biomek FX. About 20 μl (ul) of TNF-α     standard (Cat No. 210-TA; R&D Systems Inc., 614 McKinley Place NE,     Minneapolis, Minn. 55413, USA) are added to column 12 of the MSD     plate to generate a standard calibration curve (about 0 to 30000     pg/ml final). About 20 μl (ul) of diluted sulfo-TAG antibody (ca. 1     μg/ml working concentration) is added to each well, and the     plates/wells are shaken at room temperature for about 2 hours.     Finally, about 90 μl (ul) of MSD Read Buffer P (diluted to 2.5 times     with distilled water) is added and the plates are read on a MSD     Sector 6000.     Data Analysis:

Data analysis is performed with ActivityBase/XC50 module (ID Business Solutions Ltd., 2 Occam Court, Surrey Research Park, Guildford, Surrey, GU2 7QB UK) or with Bioassay (Cambridgesoft 1 Signet Court Swann's Road, Cambridge, CB5 8LA, UK). Data are normalized and expressed as % inhibition using the formula 100*((U−C1)/(C2−C1)) where U is the unknown value, C1 is the average of the high signal (0%) control wells (column 10), and C2 is the average of the low signal (100%) control wells (column 11). Curve fitting is performed with the following equation: y=A+((B−A)/(1+(10^x/10^C)^D)), where A is the minimum response, B is the maximum response, C is the log10(IC₅₀), and D is the Hill slope. The XC50 module automatically constrains A, B or A and B if an acceptable unconstrained fit cannot be achieved. QC criteria are applied and fits are rejected where A<−40 or >30, B<80 or >140 or the ratio of upper and lower confidence limits on C>10. The results for each compound are recorded as pIC₅₀ values (−C in the above equation).

Compounds are considered active in this assay if they demonstrated a pIC₅₀ of greater than 5 up to a pIC₅₀ of 10 or greater, and were screened at concentrations up to 10 uM. Representative compounds of Formula (I) as described in Examples 190-224, 226-260, 262-272, 274-282, 284-310, 313-323, 325-326, and 328-433 were tested in the above assay and found to be the most active.

In Vivo Biological Assays

The in vitro enzymatic PDE4B inhibition assay(s) described herein, or generally similar or generally analogous assays should be regarded as being the primary test(s) of biological activity. However, additional in vivo biological tests which are not an essential measure of activity, efficacy or side-effects but may be used for further characterization are described below.

LPS-Induced Pulmonary Neutrophilia in Rats: Effect of i.t. Administered PDE4 Inhibitors

Pulmonary neutrophil influx is thought to be a significant component to the family of pulmonary diseases like chronic obstructive pulmonary disease (COPD) which can involve chronic bronchitis and/or emphysema (G. F. Filley, Chest. 2000; 117(5); 251 s-260s). The purpose of this neutrophilia model is to study the potentially anti-inflammatory effects in vivo of orally administered PDE4 inhibitors on neutrophilia induced by inhalation of aerosolized lipopolysaccharide (LPS), modeling the neutrophil inflammatory component(s) of COPD. See the literature section below for scientific background.

For initial screening purposes, male Lewis rats (Charles River, Raleigh, N.C., USA) weighing approximately 280-400 grams are pretreated with a single intratracheal dose (200 μl) of either 300 μg/kg, or 30 μg/kg, of the test compound suspended in 0.5% Tween 80 (Sigma-Aldrich, St Louis, Mo., USA) in phosphate buffered saline or vehicle only. Secondarily, dose response curves may be generated using intratracheal doses of 300, 30 and 10 μg/kg, again administered in 0.5% Tween 80 (Sigma-Aldrich, St Louis, Mo., USA) in phosphate buffered saline (200 μl per rat, 30 minutes prior to LPS exposure. After a predetermined pretreatment time, the rats are exposed to aerosolized LPS (Serotype E. Coli 026:B6 prepared by trichloroacetic acid extraction, Sigma-Aldrich, St Louis, Mo., USA), generated from a nebulizer containing a 100 μg/mL LPS solution. Rats are exposed to the LPS aerosol at a rate of ca. 4 L/min for. 20 minutes. LPS exposure is carried out in a closed chamber with internal dimensions of roughly 45 cm length×24 cm width×20 cm height. The nebulizer and exposure chamber are contained in a certified fume hood. At about 4 hours-post LPS exposure the rats are euthanized by overdose with pentobarbital at 90 mg/kg, administered intraperitoneally. Bronchoalveolar lavage (BAL) is performed through a 14 gauge blunt needle into the exposed trachea. Five, 5 ml washes are performed to collect a total of 25 ml of BAL fluid. Total cell counts and leukocyte differentials are performed on the BAL fluids in order to calculate neutrophil influx into the lung. For single dose experiments, percent inhibition of neutrophil number, neutrophil percent, or both may be calculated and reported for that specific dose. For the secondary dose response studies, percent neutrophil inhibitions of either neutrophil number or neutrophil percent at each dose (cf. vehicle) may be used to calculate a sigmoidal dose-response curve (variable slope) usually using Prism Graph-Pad. The dose-response curve may also be used to calculate an ED50 value (in mg per kg of body weight) for inhibition by the test compounds of the LPS-induced neutrophilia.

Various literature references include, but are not limited to:

Filley G. F. Comparison of the structural and inflammatory features of COPD and asthma.

Chest. 2000; 117(5) 251s-260s.

Howell RE, Jenkins LP, Fielding LE, and Grimes D. Inhibition of antigen-induced pulmonary eosinophilia and neutrophilia by selective inhibitors of phosphodiesterase types 3 and 4 in brown Norway rats. Pulmonary Pharmacology. 1995; 8: 83-89.

Spond J, Chapman R, Fine J, Jones H, Kreutner W, Kung TT, Minnicozzi M. Comparison of PDE 4 inhibitors, Rolipram and SB 207499 (Ariflo™), in a rat model of pulmonary neutrophilia. Pulmonary Pharmacology and Therapeutics. 2001; 14: 157-164.

Underwood DC, Osborn RR, Bochnowicz S, Webb EF, Rieman DJ, Lee JC, Romanic AM, Adams JL, Hay DWP, and Griswold DE. SB 239063, a p38 MAPK inhibitor, reduces neutrophilia, inflammatory cytokines, MMP-9, and fibrosis in lung. Am J Physiol Lung Cell Mol Physiol. 2000; 279: L895-L902.

Examples are listed as producing “significant” inhibition if the test compound demonstrated significant (p<0.05, using a two tailed distribution and two sample equal variance students T test performed in Microsoft Excel) inhibition of either neutrophil number, neutrophil percent, or both, when dosed at either 300 or 30 μg/kg, 30 minutes prior to LPS aerosol exposure.

For purposes herein:

pIC₅₀ IC₅₀ (nM) IC₅₀ (μM) 4.00 100,000.0 100 5.00 100,000.0 10 6.00 1,000.0 1 7.00 100.0 0.1 8.00 10.0 0.01 9.00 1.0 0.001 10.00 0.1 0.0001 Methods of Manufacture

The compounds of this invention may be made by a variety of methods, including standard chemistry. Any previously defined variable will continue to have the previously defined meaning unless otherwise indicated. Illustrative general synthetic methods are set out below and then specific compounds of the invention are prepared in the working Examples. For purposes herein, the compounds in the Schemes are shown generically with the formula terms, and LINK representing an alkyl linker. Where appropriate additional substituent groups as defined within the scheme, e.g. L is a leaving group, P represents a protecting group, etc.

Compounds of formula (IX) shown below, wherein R¹, R² and R³ are as defined herein and R⁴ represents hydrogen, can be prepared by hydrogenation of an azide compound of formula (XI), wherein R¹, R² and R³ are as defined herein, in the presence of a suitable catalyst such as a palladium catalyst, e.g. palladium on carbon, in a suitable solvent such as ethanol, e.g. at a suitable temperature such as room temperature:

Compounds of formula (XI), wherein R¹, R² and R³ are as defined herein, may be prepared from compounds of formula (XII), wherein R¹, R² and R³ are as defined herein and wherein X⁶ is a leaving group such as a halogen atom, mesylate (methanesulfonate), tosylate (p-toluenesulfonate), or triflate (trifluoromethanesulfonate) (suitably a halogen atom such as a chlorine atom).

For example the compounds of formula (XII), e.g. wherein X⁶ is Cl, can be reacted with an azide salt such as sodium, lithium or potassium azide, in a suitable solvent such as dimethylsulfoxide such as dry DMSO, e.g. at a suitable temperature such as room temperature, to give compounds of formula (XI).

Compounds of formula (XII), wherein R¹, R² and R³ and X⁶ are as defined herein, can be prepared by reaction of compounds of formula (XIII), wherein R¹, R² and R³ are as defined herein, with a suitable reagent such as thionyl chloride (for when X⁶ is Cl), oxalyl chloride (for when X⁶ is Cl), methanesulfonyl chloride (for when X⁶ is mesylate), or para-toluenesulfonyl chloride (for when X⁶ is tosylate), preferably thionyl chloride. Suitable conditions, for when X⁶ is Cl, include reacting with thionyl chloride in a suitable non-aqueous (e.g. anhydrous) aprotic organic solvent such as toluene, e.g. with heating to ca. 60-90° C. for example ca. 85° C. Alternative conditions include reacting compounds of formula (XIII) with thionyl chloride and methanesulfonic acid in a suitable non-aqueous (e.g. anhydrous) aprotic organic solvent such as dichloromethane, e.g. at a suitable temperature such as room temperature.

Alternatively, compounds of formula (XI) wherein R¹, R² and R³ are as defined herein can be prepared directly from compounds of formula (XIII) wherein R¹, R² and R³ are as defined herein. For example, compounds of formula (XI) may be prepared by reacting compounds of formula (XIII) with an azide salt, e.g. sodium azide, in the presence of a halogenating agent such as carbon tetrabromide and a phosphine such as triphenylphosphine under suitable conditions, such as N,N-dimethylformamide, e.g. at a suitable temperature such as between 0° C. and room temperature (see e.g Toyota et. al. Journal of Organic Chemistry (2000), 65(21), 7110-7113).

This route, (XIII) to (XI) directly, may be suitable for where R³ is a urea-containing group [such as a N-aminocarbonyl-piperidinyl or N-aminocarbonyl-pyrrolidinyl group within sub-formula (bb) or (aa), because it is noted that these R³ urea-containing groups may not be tolerant of thionyl chloride which may be used in converting (XIII) to (XII) wherein X⁶ is Cl and onward to (XI).

In another alternative embodiment of particular interest, an amine compound of formula (IX) or a salt thereof (e.g. HCl salt thereof), wherein R¹, R² and R³ are as defined herein and R⁴ is as defined herein (in particular where R⁴ is a hydrogen atom), may be prepared directly from a compound of formula (XII) or a salt thereof, wherein R¹, R² and R³ and X⁶ are as defined herein, without first converting to an azide compound of formula (XI). For example, in compound (XII), X⁶ can in particular be a chlorine atom. When X⁶ is a chlorine atom, a benzenesulfonate salt of the compound of formula (XII) can for example be used, in particular when R¹ and R² are ethyl and when R³ is, for instance, a tetrahydro-2H-pyran-4-yl.

The reaction of the compound (XII) or the salt thereof to the amine compound (IX) or the salt thereof may optionally be carried out under suitable conditions, for example by reaction of a compound of formula (XII) or a salt thereof with an aminating agent. When R⁴ represents a hydrogen atom, and optionally for example when X⁶ is a chlorine atom, a suitable aminating agent may be used, e.g. an alkali-metal hexamethyldisilazide such as lithium hexamethyldisilazide, sodium hexamethyldisilazide or potassium hexamethyldisilazide (in particular lithium hexamethyldisilazide, e.g. with slow mixing/addition), in a suitable non-aqueous non-alcohol (aprotic) organic solvent (e.g. anhydrous solvent) such as tetrahydrofuran, for example at a suitable temperature such as about 25 to about 50° C., for example ca. 30-45° C. or ca. 30-40° C. The reaction with the suitable aminating agent (e.g. with the alkali-metal hexamethyldisilazide) is suitably followed by treatment with an aqueous acid such as aqueous hydrochloric acid (e.g. 2-10M, e.g. about 5M), for example at a suitable temperature such as from 0° C. to room temperature, for example at 5-15° C. or ca. 10° C. Optionally, extraction of an organic solution of (IX) or a salt thereof with aqueous base such as conc. (e.g. 32% w/w) NaOH solution, can be used to form the amine compound (IX) as the “free base”. Optionally, a mono-acid-addition salt, e.g. monohydrochloride, of the amine (IX) can be formed by converting the “free base” amine compound (IX) with about 1 equivalent (e.g. 1.03 equiv.) of a suitable acid such as HCl (e.g. aqueous hydrochloric acid such as ca. 36% w/w aq. HCl).

In a simplified embodiment of the process from compound (XII) or a salt thereof to an amine compound (IX) or a salt thereof, when X⁶ is a chlorine atom in the compound of formula (XII) and when R⁴ is a hydrogen atom in the compound of formula (IX), the precursor alcohol compound of formula (XIII) or a salt thereof is converted into the amine of formula (IX) or a salt thereof, via the compound of formula (XII) or a salt thereof, without substantially purifying and/or without substantially isolating the compound of formula (XII) or the salt thereof wherein X⁶ is a chlorine atom. In this embodiment, the compound of formula (XII) or the salt thereof wherein X⁶ is a chlorine atom can for example be in the form of the benzenesulfonate salt, in particular when R¹ and R² are ethyl and when R³ is for instance, a tetrahydro-2H-pyran-4-yl:

Compounds of formula (XIII) below, wherein R¹, R² and R³ are as defined herein, can be prepared by reaction of compounds of formula (XIV), wherein R¹, R² and R³ are as defined herein, and wherein X⁷ is an alkyl group such as a C₁₋₆ or C₁₋₄ alkyl (e.g. straight-chain alkyl) group e.g. in particular ethyl, with a suitable reducing agent in a suitable solvent, e.g. at a suitable temperature. One suitable reducing agent is lithium borohydride, in which case: a suitable solvent can be a mixture of tetrahydrofuran (e.g. dry) and methanol (e.g. dry) optionally also with toluene (e.g. dry), or THF, or methanol, and/or a suitable reaction temperature can be from room temperature to the reflux temperature, e.g. about 50 to about 75° C., e.g. about 60 to about 70° C., e.g. 63-69° C. or 64-68° C. Another reducing agent is di-iso-butylaluminium hydride (e.g. solution in toluene), in which case: a suitable solvent is dichloromethane and/or toluene, and/or a suitable reaction temperature can be about 0° C.

Compounds of formula (XIV), wherein R¹, R² and R³ and X⁷ are as defined herein, may be prepared by reaction of a compound of formula (XV) with an amine of formula R³NH₂, for example generally according to the method described by Yu et. al. in J. Med. Chem., 2001, 44, 1025-1027. The reaction is preferably carried out in the presence of a base such as triethylamine or N,N-diisopropylethylamine, and/or in an organic solvent such as ethanol, dioxane, 1-methyl-2-pyrrolidinone (NMP) or acetonitrile. The reaction may require heating e.g. to ca. 60-180° C., for example at 115° C.:

When R³ is a N-aminocarbonyl-piperidinyl or N-aminocarbonyl-pyrrolidinyl group the compound of formula (XIV) can be prepared by reacting a compound of formula (XIVa), below, wherein R¹, R² and X⁷ are as defined herein and n⁴=0 or 1, or a salt thereof (e.g. a hydrochloride salt thereof) with a urea-forming reagent capable of converting the (4-piperidinyl)amino or (3-pyrrolidinyl)amino group in the compound of formula (XIVa) into a [(1-aminocarbonyl)-4-piperidinyl]amino group or [(1-aminocarbonyl)-3-pyrrolidinyl]amino group as in formula (XIV) respectively:

The urea-forming reagent may be benzyl isocyanate (followed later by debenzylation e.g. reductive debenzylation), or preferably the urea-forming reagent is tri(C₁₋₄ alkyl)silyl isocyanate such as a tri(C₁₋₂alkyl)silyl isocyanate, preferably trimethylsilyl isocyanate. The conversion of the compound (XIVa) or salt thereof to the compound (XIV) may be carried out in the presence of a suitable base such as N,N-diisopropylethylamine, in a suitable solvent such as dichloromethane or chloroform, at a suitable temperature such as at room temperature or at the reflux temperature of the solvent.

Compound (XIVa), wherein R¹, R², X⁷ and n⁴ are as defined herein, or the salt thereof can be prepared from compound (XIVb) below, wherein R¹, R², X⁷ and n⁴ are as defined herein and Prot is a suitable nitrogen protecting group such as (tert-butyloxy)carbonyl, by removal of the nitrogen protecting group. For example, removal of the (tert-butyloxy)carbonyl group can be effected under suitable acidic conditions, such as with hydrogen chloride (e.g. 4M) in a suitable solvent such as 1,4-dioxane:

Compound (XIVb), wherein R¹, R², and n⁴ are as defined herein, X⁷ is ethyl and Prot is (tert-butyloxy)carbonyl, can be prepared by reaction of a compound of formula (XV), wherein R¹ and R² are as defined herein and X⁷=ethyl, with 1,1-dimethylethyl 4-amino-1-piperidinecarboxylate (e.g. commercially available from AstaTech, Philadelphia, USA) or 1,1-dimethylethyl 3-amino-1-pyrrolidinecarboxylate (e.g. commercially available from Aldrich). The reaction is optionally carried out in the presence of a base such as triethylamine or N,N-diisopropylethylamine, optionally in a suitable organic solvent such as acetonitrile, at a suitable temperature such as 60-100° C. (e.g. 80-90° C.).

Compounds of formula (XV), wherein R¹, R², and X⁷ are as defined herein can be prepared by reaction of compounds of formula (XVI), wherein R¹ is as defined herein, with a dialkyl (1-chloroalkylidene)propanedioate, for example a diethyl (1-chloroalkylidene)-propanedioate of formula (XVII) (wherein R² and X⁷ are as defined herein), followed by reaction with phosphorous oxychloride. Suitable conditions for reaction of compounds of formula (XVI) with a dialkyl (1-chloroalkylidene)propanedioate of formula (XVII) include heating in a suitable solvent such as toluene, in the presence of a suitable base such as triethylamine, at a suitable temperature such as the reflux temperature of the solvent. Suitable conditions for the reaction of the intermediate with phosphorous oxychloride include heating at the reflux temperature of phosphorous oxychloride.

Compounds of formula (XVII), wherein R² and X⁷ are as defined herein, may be prepared by reaction of compounds of formula (XVIII), wherein R² and X⁷ are as defined herein, with phosphorus oxychloride in the presence of a suitable base such as tributylamine, at a suitable temperature such as 80-130° C., for example ca. 100-120° C.

Compounds of formula (XVIII), wherein R² and X⁷ are as defined herein, may be prepared by reaction of a dialkyl malonate of formula (XIX), wherein X⁷ is as defined herein, with magnesium chloride and a suitable base such as triethylamine, in a suitable solvent such as acetonitrile, at a suitable temperature such as 5-10° C., followed by addition of an acid chloride of formula (XX), for example propanoyl chloride, at a suitable temperature such as between 10° C. and room temperature.

Compounds of formulae (XIX) and (XX) are either known compounds or may be prepared by conventional means. For example compounds of formulae (XIX) and (XX) where X⁷ and R² respectively represent ethyl are available from Aldrich.

Compounds of formula (XV), wherein R¹, R² and X⁷ are as defined herein, may alternatively be prepared by reaction of a compound of formula (XVI), wherein R¹ is as defined herein, with compounds of formula (XXI), wherein R² and X⁷ are as defined herein, with heating, followed by reaction with phosphorous oxychloride, again with heating (see Yu et. al. in J. Med. Chem., 2001, 44, 1025-1027). Compounds of formula (XXI) can for example be diethyl [(ethyloxy)methylidene]propanedioate (wherein R² is H and X⁷ is Et, available from Aldrich) or diethyl [1-(ethyloxy)ethylidene]propanedioate (wherein R² is Me and X⁷ is Et, see Eur. Pat. Appl. (1991), EP 413918 A2).

Where the desired amino pyrazole of formula (XVI) is not commercially available, preparation can be achieved using methods described by Dorgan et. al. in J. Chem. Soc., Perkin Trans. 1, (4), 938-42; 1980, by reaction of 3-hydrazinopropanenitrile (available from Lancaster Synthesis) with a suitable aldehyde of formula R⁴⁰CHO in a suitable solvent such as ethanol, with heating, followed by reduction with, for example sodium in a suitable solvent such as t-butanol. R⁴⁰ should be chosen so as to contain one less carbon atom than R¹, for example R⁴⁰=methyl will afford R¹=ethyl.

In an alternative embodiment of Process A, the 4-chloro substituent in the compound of formula (XV) can be replaced by another halogen atom, such as a bromine atom, or by another suitable leaving group which is displaceable by an amine of formula R³NH₂. The leaving group can, for example, be an alkoxy group —OR³⁵ such as —OC₁₋₄alkyl (in particular —OEt) or a group —O—S(O)₂—R³⁷, wherein R³⁷ is methyl, CF₃, or phenyl or 4-methyl-phenyl. The reaction may be carried out with or without solvent and may require heating.

Compounds of formula (XI), wherein R¹ and R² are as defined herein and R³ represents the N-aminocarbonyl-piperidinyl or N-aminocarbonyl-pyrrolidinyl group of sub-formula (bb) or (aa), may alternatively be prepared from compounds for formula XXXVIII, wherein R¹ and R² are as defined herein, n³ is 0 or 1, and Proc represents a suitable protecting group such as tert-butoxycarbonyl. Suitable conditions include treatment suitable acidic conditions such as hydrogen chloride in a suitable solvent such as 1,4-dioxane at a suitable temperature such as room temperature.

Compounds for formula XXXVIII, wherein R¹ and R², n⁴ and Proc are as defined herein, may be prepared from compounds for formula XXXIX, wherein R¹ and R², n³ and Proc are as defined herein. Suitable conditions include reaction of compounds of formula XXXIX with an azide such as sodium azide and a halogenating agent such as carbon tetrabromide, in the presence of a suitable phosphine such as triphenylphosphine, in a suitable solvent such as N,N-dimethylformamide, at a suitable temperature such as between 0° C. and room temperature.

Compounds of formula (XXXIX), wherein R¹ and R², n⁴ and Proc are as defined herein, may be prepared from compounds of formula (XL), wherein R¹ and R², n⁴, Proc and X⁷ are as defined herein, by reduction with a suitable reducing agent such as lithium borohydride, in a suitable solvent such as a mixture of tetrahydrofuran and methanol, at a suitable temperature such as at the reflux temperature of the solvent.

Compounds of formula (XL), wherein R¹ and R², n⁴, Proc and X⁷ are as defined herein, may be prepared from compounds of formula (XV), wherein R¹, R², and X⁷ are as defined herein, by reaction of a compound of formula (XV) with an amine of formula (XLI), wherein Proc and n⁴ are as defined herein. The reaction is preferably carried out in the presence of a base such as triethylamine or N,N-diisopropylethylamine, and/or in an organic solvent such as ethanol, dioxane, 1-methyl-2-pyrrolidinone (NMP) or acetonitrile. The reaction may require heating e.g. to ca. 60-180° C., for example at 120° C.:

Compounds of formula (XIV) wherein R² represents fluoroalkyl (for example trifluoromethyl) may be prepared according to the following Scheme A and followed by subsequent steps such as those described in other schemes herein:

Compounds of formula (IX) wherein R⁴ represents methyl or ethyl may be prepared according to the following scheme, wherein R^(n) represents H when R⁴ is methyl and R^(n) represents methyl when R⁴ is ethyl:

The following schemes are directed to the preparation of preparing compounds of Formula (I) as defined herein.

Scheme 1 above describes a general synthesis for compounds of the formula 1-4. Compound 1-1 is coupled to an appropriately protected bis-carboxylic acid (Z═CO₂H; Z₁═CO₂H), bis-sulfonic acid (Z═SO₂H; Z₁═SO₂H) or carboxylic, sulfonic acid (Z═CO₂H; Z₁═SO₂H or Z═SO₂H; Z₁═CO₂H) to give 1-2. When Z═CO₂H, the suitable protected linker is treated with a coupling agent such as DCC, EDC, HATU, HBTU, with or without the addition of HOBt, in the presence of a tertiary amine such as triethyl amine or diisopropyl ethyl amine in a solvent such as methylene chloride or DMF. When Z═SO₂H, the sulfonic acid is first converted to the corresponding sulfonyl chloride with a reagent such as thionylchloride or POCl₃. The resulting sulfonyl chloride (Z═SO₂C1) is then added to 1-1 in the presence of a tertiary amine such as triethyl amine or diisopropyl ethyl amine in a solvent such as methylene chloride to give 1-2. Intermediate 1-2 is then de-protected using methods dependent on the nature of the protecting group. For example, when 1-2 is protected as a methyl or ethyl ester (Z₁═CO₂Me or Z₁═CO₂Et), 1-2 is treated with solution of an aqueous base, e.g. NaOH, LiOH, in an organic solvent such as methanol, ethanol or dioxane. The resulting carboxylic acid (Z₁═CO₂H) and a suitable protected, where necessary, Ar₁-Ar₂ amine 1-3 is treated with a coupling agent such as DCC, EDC, HATU, HBTU, with or without the addition of HOBt, in the presence of a tertiary amine such as triethyl amine or diisopropyl ethyl amine in a solvent such as methylene chloride or DMF. For example, a suitable protecting group is needed when R₆ contains a primary or secondary amine. The resulting intermediate 1-4 is then deprotected in a method defined by the nature of the protecting group used. In the case of an acid labile amine protecting group like Boc, deprotection can be achieved using a strong acid such as TFA in a solvent such as dichloromethane to give 1-5.

Scheme 2 above describes a synthesis of the compounds where Z and Z₁═CO₂H. This procedure is particularly amenable to array format. A mixture of compound 2-1, a bis-carboxylic acid 2-2 (Z═CO₂H; Z₁═CO₂H), and a suitable protected, where necessary, Ar₁-Ar₂ amine 2-3 is treated with a coupling agent such as DCC, EDC, HATU, HBTU, with or without the addition of HOBt, in the presence of a tertiary amine such as triethyl amine or diisopropyl ethyl amine in a solvent such as methylene chloride or DMF. For example, a suitable protecting group is needed when R₆ contains a primary or secondary amine. The resulting intermediate 2-4 is then deprotected in a method defined by the nature of the protecting group used. In the case of an acid labile amine protecting group like Boc, deprotection can be achieved using a strong acid such as TFA in a solvent such as dichloromethane to give 2-5.

Scheme 3 above describes an alternate synthesis for compounds of the formula in compound 3-5 (wherein the P on the R6 designates a protected functionality). Intermediate 3-1 is deprotected using methods dependent on the nature of the protecting group. For example, when 3-1 is protected as a methyl or ethyl ester (Z₁═CO₂Me or Z₁═CO₂Et), 3-1 is treated with solution of an aqueous, e.g. NaOH, LiOH in an organic solvent such as methanol, ethanol or dioxane. The resulting carboxylic acid (Z₁═CO₂H) and Ar₁ amine 3-2 with the Ar₁ substituted with a bromine or iodine (XL=Br, I) is treated with a coupling agent such as DCC, EDC, HATU, HBTU, with or without the addition of HOBt, in the presence of a tertiary amine such as triethyl amine or diisopropyl ethyl amine in a solvent such as methylene chloride or DMF to give intermediate 3-3. Suzuki coupling of 3-3 with a suitably protected boronic acid or boronic ester 3-4 (R=boronic acid, boronic ester) in the presence of a palladium catalyst, such as Pd(PPh₃)₄, or Pd(OAc)₂/PPh₃ gives intermediate 3-5. Alternately, Stille coupling of 3-3 with a suitably protected trialkyl tin 3-4 (R=trialkyl tin) in the presence of a palladium catalyst, such as Pd(PPh₃)₄, or Pd(OAc)₂/PPh₃ gives intermediate 3-5. The resulting intermediate 3-5 is then deprotected in a method defined by the nature of the protecting group used. In the case of an acid labile amine protecting group like Boc, deprotection can be achieved using a strong acid such as TFA in a solvent such as dichloromethane to give 3-6.

Scheme 4 above describes an alternate synthesis for compounds of the general formula shown in compound 4-5 (wherein the P on the R6 designates a protected functionality). Suzuki coupling of intermediate 4-1 with a boronic acid or boronic ester aldehyde 4-2 (R=boronic acid, boronic ester) in the presence of a palladium catalyst, such as Pd(PPh₃)₄, or Pd(OAc)₂/PPh₃ gives intermediate 4-3. Alternately, Stille coupling of 4-2 with a trialkyl tin aldehyde (R=trialkyl tin) in the presence of a palladium catalyst, such as Pd(PPh₃)₄, or Pd(OAc)₂/PPh₃ gives intermediate 4-3. Reductive amination of the resulting intermediate 4-3 with a suitably protected R₆—H using a reducing reagent such as NaBH₃CN in methanol or NaBH(OAc)₃ in dichloroethane or DMF gives intermediate 4-4. Intermediate 4-4 is then deprotected in a method defined by the nature of the protecting group used. In the case of an acid labile amine protecting group like Boc, deprotection can be achieved using a strong acid such as TFA in a solvent such as dichloromethane to give 4-5.

Experimentals Section

The invention will now be described by reference to the following examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention.

General Procedures

All temperatures are given in degrees Celsius, all solvents are highest available purity and all reactions run under anhydrous conditions in an argon (Ar) or nitrogen (N₂) atmosphere where necessary.

Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates were used for thin layer chromatography. Both flash and gravity chromatography were carried out on E. Merck Kieselgel 60 (230-400 mesh) silica gel. The CombiFlash system used for purification in this application was purchased from Isco, Inc. CombiFlash purification was carried out using prepacked silica gel columns, a detector with UV wavelength at 254 nm and a variety of solvents or solvent combinations. Preparative hplc was performed using a Gilson Preparative System with variable wavelength UV detection or an Agilent Mass Directed AutoPrep (MDAP) system with both mass and variable wavelength UV detection. A variety of reverse phase columns, e.g., Luna 5 u C18(2) 100 A, SunFire™ C18, XBridge™ C18 were used in the purification with the choice of column support dependent upon the conditions used in the purification. The compounds are eluted using a gradient of acetonitrile and water. Neutral conditions used an acetonitrile and water gradient with no additional modifier, acidic conditions used an acid modifier, usually 0.1% TFA (added to both the acetonitrile and water) and basic conditions used a basic modifier, usually 0.1% NH₄OH (added to the water). Analytical hplc was run using an Agilent system with variable wavelength UV detection using reverse phase chromatography with an acetonitrile and water gradient with a 0.05 or 0.1% TFA modifier (added to each solvent). LC-MS was determined using either a PE Sciex Single Quadrupole LC/MS API-150 or a Waters. The compound is analyzed using a reverse phase column, e.g., Thermo Aquasil/Aquasil C18, Acquity HPLC C18, Thermo Hypersil Gold eluted using an acetonitrile and water gradient with a low percentage of an acid modifier such as 0.02% TFA or 0.1% formic acid.

Nuclear magnetic resonance spectra were recorded at 400 MHz using a Bruker AC 400 or Brucker DPX400 spectrometer. CDCl₃ is deuteriochloroform, DMSO-D₆ is hexadeuteriodimethylsulfoxide, and CD₃OD is tetradeuteriomethanol. Chemical shifts are reported in parts per million (δ) downfield from the internal standard tetramethylsilane (TMS) or calibrated to the residual proton signal in the NMR solvent (e.g., CHCl₃ in CDCl₃). Abbreviations for NMR data are as follows: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, dd=doublet of doublets, dt=doublet of triplets, app=apparent, br=broad. J indicates the NMR coupling constant measured in Hertz. Melting points were determined using a Electrothermal 9100 apparatus (Electrothermal Engineering Ltd.).

Heating of reaction mixtures with microwave irradiation was carried out on a Smith Creator (purchased from Personal Chemistry, Forboro, Mass., now owned by Biotage), an Emrys Optimizer (purchased from Personal Chemistry) or an Explorer (purchased from CEM, Matthews, N.C.) microwave.

Cartridges or columns containing polymer based functional groups (acid, base, metal chelators, etc) can be used as part of compound workup. The “amine” columns or cartridges are used to neutralize or basify acidic reaction mixtures or products. These include NH2 Aminopropyl SPE-ed SPE Cartridges available from Applied Separations and diethylamino SPE cartridges available from United Chemical Technologies, Inc.

Abbreviations are listed in the table below. All other abbreviations are as described in the ACS Style Guide (American Chemical Society, Washington, D.C., 1986).

Table of Abbreviations DMAP: 4-(Dimethylamino)pyridine SPE: Solid phase extraction DCM: Dichloromethane m-CPBA: 3-Chlorobenzene- DMF: N,N-Dimethylformamide carboperoxoic acid dppf: 1,1′-Bis(diphenylphosphino)- MDAP: Mass directed auto ferrocene preparation DMSO: Dimethylsulfoxide NIS: N-Iodosuccinimide DIPEA: N,N-Diisopropylethylamine HATU: O-(7-Azabenzotriazol-1- DSC: differential scanning calorimetry yl)-N,N,N′,N′-tetramethyluronium EtOAc: Ethyl acetate hexafluorophosphate EDC: 1-(3-Dimethylaminopropyl)-3- HBTU: O-Benzotriazol-1-yl- ethylcarbodiimide hydrochloride N,N,N′,N′-tetramethyluronium TFA: Trifluoroacetic anhydride hexafluorophosphate M: molar HOBT: 1-Hydoxybenzotriazole L: liters hydrate mL: milliliters IPA: isopropyl alcohol g: grams THF: Tetrahydrofuran mg: milligrams mol: moles h: hours mmol: millimoles NMP: 1-methyl-2-pyrrolidinone Satd: saturated TEA: triethylamine eq: equivalents CDI: carbonyldiimidazole min: minutes TSBCl: tert-butyldimethylsilyl mp: melting point chloride rt: room temperature Aq or aq: aqueous NBS: N-bromosuccinimide BPO: benzoyl peroxide

EXAMPLES Intermediate Compounds Example A N-[(3-Bromophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

The 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic acid (931 mg, 2 mmol) was partially dissolved in dichloromethane (50 mL), and HBTU (758 mg, 2.000 mmol) was added followed sequentially by [(3-bromophenyl)methyl]amine hydrochloride (464 mg, 2.000 mmol), and TEA (0.558 mL, 4.00 mmol). The mixture was allowed to stir under argon overnight at room temperature. The solvent was removed on a rotavap and the residue was taken up in EtOAc (100 mL) and washed 3× with water (50 mL) (emulsion, a few drops of 1N NaOH helps break emulsion), 1× with brine, dried over anhydrous Na₂SO₄, filtered and evaporated to give the crude residue. The residue was taken up in methylene chloride and absorbed on Isolute® Sorbent and purified by Combiflash using an 80 g silica column eluted with 0-10% DCM/MeOH. The product fractions were combined and concentrated to give N-[(3-bromophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (962 mg, 1.476 mmol, 73.8% yield) as a white solid. LC-MS m/z 633.6 (M+H)⁺, 0.97 min (ret time); mp 116-118° C.

Example B N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide

N-[(3-Bromophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (0.825 g, 1.302 mmol), (3-formylphenyl)boronic acid (0.195 g, 1.302 mmol), potassium carbonate (0.540 g, 3.91 mmol), and Pd(Ph₃P)₄ (0.075 g, 0.065 mmol) were divided into 3 portions and added to three 2-5 mL Biotage microwave vials in 1,4-dioxane (3 mL) and water (1 mL). The vials were capped and the mixtures were microwaved at normal power in the Emrys Optimizer at 100° C. for 15 min. The combined crude product was partitioned between EtOAc (75 mL) and water (20 mL). The phases were separated, and the organic phase was washed with water (2×20 mL), brine (20 mL), dried over anhydrous Na₂SO₄ filtered and evaporated to give the crude residue. The residue was taken up in DCM and absorbed on Isolute® Sorbent and purified by Combiflash using an 80 g silica column eluted with 0-10% DCM/MeOH. The product fractions were combined and concentrated to give (N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (584.1 mg, 0.875 mmol, 67.2% yield)) as a white solid. LC-MS m/z 659.6 (M+H)⁺, 1.01 min (ret time); mp 109-112° C.

Example C N-[(3-Bromo-4-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

The 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic acid (931 mg, 2 mmol) was partially dissolved in dichloromethane (50 mL), and HBTU (758 mg, 2.000 mmol) was added followed sequentially by [(3-bromo-4-fluorophenyl)methyl]amine hydrochloride (481 mg, 2.000 mmol), and TEA (0.836 mL, 6.00 mmol). The mixture was allowed to stir under argon overnight at room temperature. The solvent was removed on a rotavap. The residue was taken up in EtOAc (100 mL) and washed 3× with water (50 mL) (emulsion, a few drops of 1N NaOH helps break emulsion), 1× with brine, dried over anhydrous Na₂SO₄, filtered and evaporated to give the crude residue. It was taken up in methylene chloride and absorbed on Isolute® Sorbent and purified by Combiflash using an 80 g silica column eluted with 0-10% DCM/MeOH. The product fractions were combined and concentrated to give N-[(3-bromo-4-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (875 mg, 1.329 mmol, 66.5% yield) as a white solid. LC-MS m/z 651.3 (M+H)⁺, 0.98 min (ret time).

Example D N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide

N-[(3-Bromo-4-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (0.849 g, 1.303 mmol), (3-formylphenyl)boronic acid (0.195 g, 1.303 mmol), potassium carbonate (0.540 g, 3.91 mmol), and Pd(Ph₃P)₄ (0.075 g, 0.065 mmol) were divided into 3 portions and added to three 2-5 mL Biotage microwave vials in 1,4-dioxane (3 mL) and water (1 mL). The vials were capped and the mixtures were microwaved at normal power in the Emrys Optimizer at 100° C. for 15 min. The combined crude product was partitioned between EtOAc (75 mL) and water (20 mL). The phases were separated, and the organic phase was washed with water (2×20 mL), brine (20 mL), dried over anhydrous Na₂SO₄ filtered and evaporated to give the crude material. It was taken up in DCM and absorbed on Isolute® Sorbent and purified by Combiflash using an 80 g silica column eluted with 0-10% DCM/MeOH. The product fractions were combined and concentrated to give (N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (596.6 mg, 0.874 mmol, 67.0% yield)) as a beige solid. LC-MS m/z 677.4 (M+H)⁺, 1.01 min (ret time); mp 107-110° C.

Example E N-[(3-Bromo-4-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

The 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic acid (931 mg, 2 mmol) was partially dissolved in the dichloromethane (50 mL), and HBTU (758 mg, 2.000 mmol) was added followed sequentially by [(3-bromo-4-chlorophenyl)methyl]amine (441 mg, 2.000 mmol), and TEA (0.558 mL, 4.00 mmol). The mixture was allowed to stir under argon overnight at room temperature. The solvent was removed on a rotavap. The residue was taken up in EtOAc (100 mL) and washed 3× with water (50 mL) (emulsion, a few drops of 1N NaOH helps break emulsion), 1× with brine. A substantial amount of solid would not dissolve in either the EtOAc or aqueous phases. This was filtered off and set aside. The organic phase was dried over anhydrous Na₂SO₄, filtered and evaporated to give N-[(3-bromo-4-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (349 mg, 0.324 mmol, 16.20% yield). The solid that had been set aside was washed with water and ethyl acetate and dried under vacuum to give another batch of N-[(3-bromo-4-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (993.7 mg, 1.488 mmol, 74.4% yield) as a white solid. LC-MS m/z 667.2 (M+H)⁺, 1.02 min (ret time); mp 212-214° C.

Example F N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

N-[(3-Bromo-4-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (868 mg, 1.3 mmol), (3-formylphenyl)boronic acid (195 mg, 1.300 mmol), potassium carbonate (539 mg, 3.90 mmol), and Pd(Ph₃P)₄ (75 mg, 0.065 mmol) were divided into 3 portions and added to three 2-5 mL Biotage microwave vials in 1,4-dioxane (3 mL) and water (1 mL). The vials were capped and the mixtures were microwaved at normal power in the Emrys Optimizer at 100° C. for 15 min. The combined crude product was partitioned between EtOAc (75 mL) and water (20 mL). The phases were separated, and the organic phase was washed with water (2×20 mL), brine (20 mL), dried over anhydrous Na₂SO₄ filtered and evaporated to give the crude residue. The residue was taken up in DCM and absorbed on Isolute® Sorbent and purified by Combiflash using an 80 g silica column eluted with 0-10% DCM/MeOH. The product fractions were combined and concentrated to give (N-[(6-chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (531.4 mg, 0.759 mmol, 58.4% yield) as a white solid. LC-MS m/z 693.4 (M+H)⁺, 1.01 min (ret time); mp 153-156° C.

Example G N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide

N-{[3-Bromo-4-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (900 mg, 1.356 mmol), (3-formylphenyl)boronic acid (203 mg, 1.356 mmol), potassium carbonate (562 mg, 4.07 mmol), and Pd(Ph₃P)₄ (78 mg, 0.068 mmol) were divided into 3 portions and added to three 2-5 mL Biotage microwave vials in 1,4-dioxane (3 mL) and water (1 mL). The vials were capped and the mixtures were microwaved at normal power in the Emrys Optimizer at 100° C. for 15 min. The combined crude product was partitioned between EtOAc (75 mL) and water (20 mL). The phases were separated and the organic phase was washed with water (2×20 mL), brine (20 mL), dried over anhydrous Na₂SO₄, filtered and evaporated to give the crude residue. It was taken up in DCM and absorbed on Isolute® Sorbent and purified by CombiFlash using an 80 g silica column eluted with 0-10% MeOH/DCM. The product fractions were combined and concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide (632 mg, 0.873 mmol, 64.3% yield) as a white solid. LC-MS m/z 689.8 (M+H)⁺, 0.95 min (ret time); mp 117-119° C.

Example H N-[(3-Bromo-4-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

The 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic acid (0.931 g, 2 mmol) was partially dissolved in the dichloromethane (50 mL), and HBTU (0.758 g, 2.000 mmol) was added followed sequentially by [(3-bromo-4-methylphenyl)methyl]amine (0.400 g, 2.000 mmol), and TEA (0.558 mL, 4.00 mmol). The mixture was allowed to stir under argon overnight at room temperature. The solvent was removed on a rotavap and the residue was taken up in EtOAc (100 mL) and washed 3× with water (50 mL) (emulsion formed, a few drops of 1N NaOH helps break emulsion), 1× with brine, dried over anhydrous Na₂SO₄, filtered and evaporated to give the crude residue. The residue was taken up in methylene chloride and absorbed on Isolute® Sorbent and purified by Combiflash using an 80 g silica column eluted with 0-10% DCM/MeOH. The product fractions were combined and concentrated to give N-[(3-bromo-4-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (1.073 g, 1.635 mmol, 82% yield) as a white solid. LC-MS m/z 647.4 (M+H)⁺, 0.98 min (ret time); mp 117-119° C.

Example I N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide

N-[(3-Bromo-4-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (840 mg, 1.297 mmol), (3-formylphenyl)boronic acid (194 mg, 1.297 mmol), potassium carbonate (538 mg, 3.89 mmol), and Pd(Ph₃P)₄ (74.9 mg, 0.065 mmol) were divided into 3 portions and added to three 2-5 mL Biotage microwave vials in 1,4-dioxane (3 mL) and water (1 mL). The vials were capped and the mixture was microwaved at normal power in the Emrys Optimizer at 100° C. for 15 min. The combined crude product was partitioned between EtOAc (75 mL) and water (20 mL). The phases were separated, and the organic washed with water (2×20 mL), brine (20 mL), dried over anhydrous Na₂SO₄ filtered and evaporated to give the crude residue. The residue was taken up in DCM and absorbed on Isolute® Sorbent and purified by Combiflash using an 80 g silica column eluted with 0-10% DCM/MeOH. The product fractions were combined and concentrated to give (N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (537 mg, 0.789 mmol, 60.8% yield)) as a white solid. LC-MS m/z 673.4 (M+H)⁺, 0.99 min (ret time); mp 117-120° C.

Example 1 Diethyl Propanoylpropanedioate

To magnesium chloride (2.96 g, 31.2 mmol) was added dry acetonitrile (5 mL) and the mixture was then cooled in ice and treated with diethyl malonate (5 g, 31.2 mmol). Once the mixture was cold, triethylamine (8.6 mL, 62.5 mmol) was added and the resulting suspension was stirred for 15 min. Propionyl chloride (2.71 mL, 31.2 mmol) was added dropwise and the mixture was stirred at 0° C. for 1.5 h and at ambient temperature for 5 h. The mixture was cooled in an ice-bath and treated with aqueous hydrochloric acid (2M, 10 mL) and the product extracted with ether. The organic phase was washed with water then brine, dried and concentrated to afford 6.31 g of a yellow oil. This was dissolved in ether and washed with aqueous hydrochloric acid (2M) then brine, dried and concentrated to afford 5.93 g of the title compound.

Example 2 Diethyl (1-chloropropylidene)propanedioate

To diethyl propanoylpropanedioate (5.93 g, 27.4 mmol) was added phosphorus oxychloride (38 mL) and tributylamine (6.5 mL, 27.4 mmol) and the mixture was heated to 115° C. for 6 h then stirred at ambient temperature for 16 h. The mixture was concentrated to dryness and the residue added cautiously to aqueous hydrochloric acid (1M, 80 mL) and extracted twice with diethyl ether. The combined organic layers were washed with aqueous hydrochloric acid (1M), water, aqueous sodium hydroxide (1M) then brine, dried and concentrated to dryness to afford 6.81 g of a brown oil. The product was purified by flash chromatography on silica (250 mL) eluting with ethyl acetate/cyclohexane @ 1:10 to afford 3.21 g of the title compound. LC-MS m/z 235, 237 (M+H)⁺, 3.30 min (ret time).

Example 3 Ethyl 4-chloro-1,6-diethyl-1H-pyrazolo[3,4-b]pyridine-5-carboxylate

To 1-ethyl-1H-pyrazol-5-amine (Aldrich, 1.52 g, 13.7 mmol) was added a solution of diethyl (1-chloropropylidene)propanedioate (3.21 g, 13.7 mmol) in toluene (40 mL) followed by triethylamine (3.78 mL, 27.3 mmol) and then heated at reflux for 6 h. The cooled mixture was concentrated to dryness and the resulting brown residue was treated with phosphorus oxychloride (25 mL, 0.274 mol) and heated to 110° C. for 17.5 h. The cooled mixture was concentrated to dryness and the residue was to water (caution, exotherm) and extracted with ethyl acetate. The aqueous phase was treated with aqueous sodium hydroxide (2M) to achieve pH 9 and extracted with additional ethyl acetate. The combined organics were washed with aqueous sodium bicarbonate, then brine, dried and concentrated to dryness to afford 3.6 g of a dark brown oil. The product was purified by flash chromatography on silica (150 mL) eluting with ethyl acetate/cyclohexane from 1:10 to 1:8 to afford 1.8 g of the title compound. LC-MS m/z 282 (M+H)⁺, 3.46 min (ret time).

Example 4 Ethyl 1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridine-5-carboxylate

To a solution of ethyl 4-chloro-1,6-diethyl-1H-pyrazolo[3,4-b]pyridine-5-carboxylate (380 g) in 1-methylpyrrolidine (3,166 mL) was added diisopropylethylamine (469.8 mL) and tetrahydro-2H-pyran-4-ylamine (163 g) and the mixture was heated at reflux for 16 h. The cooled mixture was treated with water (12 L) and extracted with ethyl acetate (6×1250 mL). The combined organics were washed with brine, dried over magnesium sulphate, filtered and concentrated to dryness to afford 520 g of a dark brown oil. The product was purified by flash chromatography on silica using ethyl acetate/cyclohexane @ 1:4-1:2 as eluant to afford 336 g of the title compound. LC-MS m/z 347 (M+H)⁺, 3.02 min (ret time).

Example 5 [1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methanol

To ethyl 1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridine-5-carboxylate (60.43 g, 174 mmol) in dry THF (300 mL) was added dry methanol (28.3 mL) followed by the addition of lithium borohydride (2M in THF, 262 mL, 523 mmol) over 30 min. The mixture was heated to reflux. After 1 h additional methanol (14.1 mL) was added. After a further 30 min additional methanol (14.1 mL) was added. After a further 30 min the mixture was cooled in an ice bath and treated with methanol (100 mL) followed (cautiously) with water (1000 mL). The mixture was stirred for 1 h and then extracted with dichloromethane (1,500 mL total). The combined organics were washed with water, then brine, dried and concentrated to dryness to afford 49.84 g of the title compound. LC-MS m/z 305 (M+H)⁺, 1.79 min (ret time).

Example 6 5-(Azidomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine

To [1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methanol (24.9 g, 82 mmol) was added thionyl chloride (90 mL, 1.23 mmol) and the mixture was heated to 80° C. under an atmosphere of nitrogen. After 2 h the mixture was cooled, concentrated and the residue azeotroped with toluene. The residue was then dissolved in a solution of sodium azide (7.98 g, 123 mmol) in DMSO (120 mL). The mixture was stirred for 16 h. The above procedure was repeated on the same scale and the 2 reactions combined for work-up. The combined DMSO mixture was partitioned between ethyl acetate and aqueous sodium bicarbonate. The aqueous phase was extracted thoroughly with ethyl acetate and the combined organics were washed with water, then brine, dried and concentrated to afford 58.9 g of a brown solid. The product was purified by flash chromatography on 1.5 kg of silica using a step gradient from 3:1 to 2:1 cyclohexane/ethyl acetate to afford 39.94 g of the title compound. LC-MS m/z 330 (M+H)⁺, 2.21 min (ret time).

Example 7 5-(Aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine

Palladium on charcoal (10%, 50% w/w water, 8 g) was treated with ethanol (200 mL) followed by a solution of 5-(azidomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (39.94 g, 121 mmol) in ethanol (1200 mL). The mixture was stirred under an atmosphere of hydrogen for 16 h. The catalyst was then removed by filtration and the filtrate removed of solvent in vacuo to reveal 41.24 g of a black oil. The product was purified by flash chromatography on 1 kg of silica using a step gradient from 5-20% methanol in dichloromethane to afford 32.66 g of the title compound. LC-MS m/z 304 (M+H)⁺, 1.71 min (ret time).

Example 8 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

A solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine, previously described in WO2007036733A1, (250 mg, 0.83 mmol) in DMF (70 mL) was treated with 3-[(methyloxy)carbonyl]benzoic acid (149 mg, 0.83 mmol), PyBOP (473 mg, 0.91 mmol) and diisopropylethylamine (0.573 mL, 3.3 mmol) and the mixture was stirred at ambient temperature under a nitrogen atmosphere for 4 h. The mixture was then concentrated to dryness and the residue treated with water (150 mL) and extracted with dichloromethane (2×100 mL). The combined organics were dried (hydrophobic frit) and concentrated to dryness to afford 383 mg of the title compound. LC-MS m/z 466 (M+H)⁺, 0.90 min (ret time).

Example 9 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

A solution of methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (460 mg, 1 mmol) in THF (30 mL) was treated with a solution of lithium hydroxide (237 mg, 10 mmol) in water (10 mL) and stirred at ambient temperature for 16 h. The residue was dissolve in minimum amount of water and the pH was adjusted to 4 by the addition of aqueous hydrochloric acid (2M). The resulting solution was concentrated to dryness to afford 446 mg of the title compound. LC-MS m/z 452 (M+H)⁺, 0.77 min (ret time).

Example 10 [5′-(Aminomethyl)-2′-fluoro-3-biphenylyl]methanol

To a solution of [3-(hydroxymethyl)phenyl]boronic acid (2 g, 13.2 mmol) in 1,4-dioxan (40 mL) was added [(3-bromo-4-fluorophenyl)methyl]amine hydrochloride (3.18 g, 13.2 mmol), potassium carbonate (9.1 g, 66 mmol) and tetrakis(triphenylphosphine)-palladium(0) (456 mg, 0.4 mmol). The mixture was split into 4×20 mL capacity microwave vials and each was treated with water (3 mL). The mixtures were each heated at 150° C. for 20 min. One sixth of the total reaction mixture was treated with water (100 mL) and extracted with ethyl acetate (2×80 mL). The combined organic phase was dried (magnesium sulphate) and concentrated to dryness and the product purified by flash chromatography on silica using 0-50% ethyl acetate/cyclohexane followed by dichloromethane/ammonia solution/methanol @ 8:1:1 as eluants. Product-containing fractions were combined and concentrated to dryness to afford 402 mg of the title compound. LC-MS m/z 463 (M+H)⁺, 0.65 min (ret time). The remaining five-sixth of the reaction mixture was worked up in the same manner and purified by flash chromatography using ethyl acetate/cyclohexane @ 1:1 followed by dichloromethane/ammonia solution/methanol @ 8:1:1 as eluants. This yielded 1.97 g of the title compound. LC-MS m/z 463 (M+H)⁺, 0.65 min (ret time).

Example 11 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (446 mg, 1 mmol) in DMF (70 mL) was added [5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methanol (230 mg, 1 mmol), PyBOP (570 mg, 1.1 mmol) and diisopropylethylamine (0.69 mL, 4 mmol) and the mixture was stirred at ambient temperature for 16 h. Additional PyBOP (518 mg, 1 mmol) and diisopropylethylamine (0.69 mL, 4 mmol) were added and the mixture stirred for an additional 2 h. The mixture was then concentrated to dryness and the residue was treated with water (400 mL) and extracted with dichloromethane (2×200 mL). The combined organics were dried (hydrophobic frit) and concentrated in vacuo and the resulting yellow oil was subjected to purification by flash chromatography (100 g silica cartridge, gradient elution from 0-100% ethyl acetate/cyclohexane) to afford 650 mg of the title compound. LC-MS m/z 665 (M+H)⁺, 0.98 min (ret time).

Example 12 2-Bromo-4-methylbenzamide

To a suspension of 2-bromo-4-methylbenzoic acid (15.0 g, 69.8 mmol) in toluene (60 mL), thionyl chloride (10.3 mL) and DMF (0.10 mL) were added and stirred at 50° C. for 3 h. After cooled to room temperature, the excess thionyl chloride was removed in vacuo. The residue was dissolved in toluene (50 mL), and the mixture was added to the solution of ammonia (25%, 60 mL). The white precipitate was filtered over Celite and dried in vacuo to afford 2-bromo-4-methylbenzamide (14.8 g, 99%).

Example 13 2-Bromo-4-methylbenzonitrile

To a suspension of 2-bromo-4-methylbenzamide (14.8 g, 69.1 mmol) in CHCl₃ was added phosphorous pentoxide (24.5 g, 172.8 mmol) and the mixture keep refluxing for 12 h. The reaction was allowed to cool to room temperature, and put into the ice water under the condition of stirring. The organic layer was separated and the aqueous layer was extracted with CHCl₃ (150 mL×2). The combined organic phase was washed with brine, and dried over Na₂SO₄. Evaporation of the solvent afforded the title compound, 2-bromo-4-methylbenzonitrile (13.3 g, 98%). ¹H NMR (400 MHz, CDCl₃): δ 2.41 (s, 3 H), 7.20 (d, J=8.0 Hz, 1 H), 7.51-7.54 (m, 2 H).

Example 14 2-Bromo-4-(bromomethyl)benzonitrile

A mixture of 2-bromo-4-methylbenzonitrile (13.3 g, 81.4 mmol), NBS (14.4 g, 84.4 mmol) and BPO (0.20 g) in CCl₄ (150 mL) was heated for 4 h at reflux. The reaction mixture was cooled to room temperature and filtered. Then the solid was washed with CCl₄ (20 mL×2) and the combined filtrate was washed successively with saturated sodium bicarbonate (50 mL), water (2×100 mL) and sodium thiosulfate (50 mL). The organic phase was dried over Na₂SO₄ and concentrated in vacuum afforded the titled compound, 2-bromo-4-(bromomethyl)benzonitrile (18.7 g, 100%).

Example 15 2-Bromo-4-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]benzonitrile

To a solution of 2-bromo-4-(bromomethyl)benzonitrile (18.0 g, 65.5 mmol) in DMF (60 mL), potassium phthalide (18.2 g, 98.2 mmol) was added, and then the mixture was stirred under reflux for 4 h. The reaction was allowed to cool to room temperature. After removing DMF under reduced pressure, the residue was dissolved in CH₂Cl₂ (200 mL), and washed with water (50 mL×2). The organic layer was dried over anhydrous sodium sulfate. After evaporation of the solvent, the residue was recrystallized from toluene and EtOH to give the product, 2-bromo-4-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]benzonitrile (13.5 g, 61%).

Example 16 4-(Aminomethyl)-2-bromobenzonitrile

To a suspension of 2-bromo-4-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]benzonitrile (8.0 g, 23.5 mmol) in EtOH (150 mL) was added hydrazine hydrate (85%, 2.76 g). The mixture was refluxed for 3 h. At room temperature 2 N HCl (60 mL) was added (pH=3), and the mixture was filtered and rinsed with water (50 mL×4). The filtrate was concentrated to about 150 mL and filtered again. After addition of NaHCO₃ to pH=9, the filtrate was extracted with CH₂Cl₂ (100 mL×3). The combined extract was washed with brine, dried over anhydrous sodium sulfate. After removing the solvent, 1N HCl in MeOH (50 mL) was added and the solvent was concentrated to afford crude material as a white solid. Recrystallization from MeOH-Et₂O yielded 4.3 g of the product, 4-(aminomethyl)-2-bromobenzonitrile (yield: 75.8%). ¹H NMR (400 MHz, D₂O): δ 4.21 (S, 2 H), 7.43 (dd, J=8.0 Hz, J=1.2 Hz, 1 H), 7.71 (d, J=8.0 Hz, 2 H); ¹³C NMR (100 MHz, D₂O): δ 42.3, 115.5, 118.0, 125.6, 128.4, 133.4, 135.5, 139.9; HPLC: retention time: 4.709 min; purity: 99.7%.

Example 17 3-Bromo-5-fluorobenzonitrile

A 250-mL round-bottom flask equipped with a magnetic stir bar was charged with 1,3-dibromo-5-fluorobenzene (7.70 g, 30.3 mmol), DMF (45 mL), pyridine (4.9 mL), and copper (I) cyanide (2.72 g, 30.3 mmol) under nitrogen. A reflux condenser was attached to the flask. The green, cloudy mixture was stirred at reflux for 3 h. Once lower Rf impurities were observed, the reaction was allowed to cool to room temperature. The reaction was quenched with 30 mL of ether, and a precipitate formed in the dark solution. The precipitate was gravity-filtered though Celite. The filtrate was rinsed three times with ether (100 mL/50 g bromide). The isolated solution was added to a separatory funnel. The organic layer was washed with a 2:1 mixture of water and concentrated ammonium hydroxide (30 mL), followed by saturated ammonium chloride solution (2×30 mL) and saturated sodium bicarbonate (30 mL). The aqueous layers were extracted with ether (3×40 mL). The organic layers were combined and dried over anhydrous sodium sulfate. The product was purified by flash column chromatography to yield 3-bromo-5-fluorobenzonitrile (2.10 g, 35%). ¹H NMR (400 MHz, CDCl₃): δ 7.62 (s, 1 H), 7.54-7.50 (m, 1 H), 7.35-7.32 (m, 1 H).

Example 18 1,1-Dimethylethyl [(3-bromo-5-fluorophenyl)methyl]carbamate

NaBH₄ (1.99 g, 52.5 mmol) was cautiously added to a solution of NiCl₂ (1.36 g, 10.5 mmol), Boc₂O (4.58 g, 21.0 mmol) and 3-bromo-5-fluorobenzonitrile (2.10 g, 10.5 mmol) in absolute ethanol (30 mL) at 0° C. (vigorous reaction with the formation of a black precipitate). Once the reaction had subsided the mixture was left to stir at room temperature for 30 min. Ethanol was removed under reduced pressure and the precipitate was dissolved in EtOAc, filtered and repeatedly washed with EtOAc. The combined organic phase was washed with saturated NaHCO₃, and dried over Na₂SO₄. After removing the solvent, the product was purified by flash column chromatography to yield 1,1-dimethylethyl [(3-bromo-5-fluorophenyl)methyl]carbamate (2.20 g, 69%). ¹H NMR (400 MHz, CDCl₃): δ 1.46 (S, 9 H), 4.28-4.32 (m, 2 H), 4.87 (br, 1 H), 6.93-7.29 (m, 3 H); ¹³C NMR (100 MHz, CDCl₃): δ 20.3, 43.6, 44.1, 79.7, 80.0, 113.0, 114.0, 117.7, 122.5, 126.0, 123.0, 141.7, 155.9, 161.5, 164.0.

Example 19 3-Bromo-2-fluorobenzoic Acid

To a stirred solution of 2,2,6,6-tetramethylpiperidine (31.1 g, 0.22 mol) in THF (200 mL) was added dropwise a solution of butyl lithium (0.22 mol) in hexane (146.7 mL) at −10° C. The mixture was stirred for 1.5 h at −10° C. and the fluoroarene (1-bromo-2-fluorobenzene) in THF (100 mL) was consecutively added to the solution at −75° C. The mixture was stirred for 2 h at −75° C. and an excess of freshly CO₂ gas bubbled into the reaction mixture. Then the reaction mixture was warmed to room temperature and stirred over night. After evaporation of the solvent, the residue was dissolved in water (150 mL), washed with diethyl ether (2×50 mL), acidified (to pH 1) and the solid was filtered off, dried under vacuum to give 24.3 g of the title compound as a white solid (yield: 55%).

Example 20 3-Bromo-2-fluorobenzamide

To a stirred solution of 3-bromo-2-fluorobenzoic acid (24.3 g, 111 mmol) in CH₂Cl₂ (100 mL) was added SOCl₂ (12.2 mL, 166 mmol). The mixture was stirred under reflux for 6 h until the solution is colorless. CH₂Cl₂ was removed under vacuum, the residue was dissolved in ethyl acetate (200 mL), then was added dropwise to NH₃H₂O (80 mL). The organic layer was wash with H₂O (50 mL×2), brine and dried Na₂SO₄, filtered and concentrated to give 23.8 g of the title compound as a white solid (98% yield).

Example 21 [(3-Bromo-2-fluorophenyl)methyl]amine

To a solution of 3-bromo-2-fluorobenzamide (3.0 g, 13.76 mmol) in THF (50 mL) was added BH₃.Me₂S (1.57 mL, 20.6 mmol) and stirred at 50° C. for 2 h (monitored by TLC). The reaction was quenched by adding HCl (20 mL, 3 N) and the result mixture was stirred for 2 h before THF was removed under vacuum. The aqueous layer was extracted with AcOEt (30 mL), and then was adjusted to pH=9.0 with NaOH (1 N). The aqueous layer was extracted with AcOEt (50 mL×2). The combined organic layers were washed with brine, dried over Na₂SO₄, filtered and concentrated to give 1.30 g of the title compound, as a colorless oil (yield: 46%). ¹H NMR (400 MHz, D₂O): δ 4.30 (S, 2 H), 7.19-7.22 (m, 1 H), 7.47 (t, J=8.8 Hz, 1 H), 7.73-7.77 (m, 1 H); ¹³C NMR (100 MHz, D₂O): δ 37.4, 108.9, 121.2, 126.2, 130.7, 135.1, 156.2, 158.6.

Example 22 1-Bromo-3-(bromomethyl)-5-methylbenzene

To a solution of 1-bromo-3,5-dimethylbenzene (25.0 g, 135.0 mmol) in CCl₄ (150 mL), 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (14.5 g, 54.0 mmol) and dibenzoyl peroxide (BPO) (0.2 g) were added and the mixture was refluxed for 7 h. After the reaction was cooled to room temperature, the precipitate was filtered though Celite, and then the solid was rinsed two times with pentane (50 mL). The combined filtrate was washed with water (50 mL), followed by saturated sodium bicarbonate (50 mL) and sodium thiosulfate (50 mL×2). The organic layer was dried over anhydrous sodium sulfate. Evaporation of the solvent afforded the compound 1-bromo-3-(bromomethyl)-5-methylbenzene (35.6 g, 99%).

Example 23 2-[(3-Bromo-5-methylphenyl)methyl]-1H-isoindole-1,3(2H)-dione

To a solution of 1-bromo-3-(bromomethyl)-5-methylbenzene (34.0 g, 128.8 mmol) in DMF (200 mL), was added potassium phthalide (28.9 g, 154.6 mmol), and the mixture was stirred under reflux for 2 h. The reaction was allowed to cool to room temperature. After the solvent was remove under reduced pressure, the residue was dissolved in CH₂Cl₂ (300 mL), and washed with water (50 mL×3). The organic layer was dried over anhydrous sodium sulfate. Evaporation of the solvent gave a white solid. The solid was recrystallized from toluene and EtOH to give the product, 2-[(3-bromo-5-methylphenyl)methyl]-1H-isoindole-1,3(2H)-dione (28.5 g, 67%). ¹H NMR (400 MHz, CDCl₃): δ 7.87-7.72 (m, 4 H), 7.36 (s, 1H), 7.23 (s, 1 H), 7.15 (s, 1 H), 4.77 (s, 2 H), 2.30 (s, 3 H).

Example 24 [(3-Bromo-5-methylphenyl)methyl]amine

To a suspension of 2-[(3-bromo-5-methylphenyl)methyl]-1H-isoindole-1,3(2H)-dione (6.5 g, 19.7 mmol) in EtOH (120 mL) was added hydrazine hydrate (85%, 2.3 g). The mixture was refluxed for 3 h. After cooling to room temperature, 2 N HCl (60 mL) was added to pH=3, and the mixture was filtered and rinsed with water (50 mL×4). The filtrate was concentrated to about 150 mL and filtered again. After addition of 2 N NaOH (60 mL) (pH=9), the filtrate was extracted with CH₂Cl₂ (50 mL×4). The combined extract was washed with brine, dried over anhydrous sodium sulfate, and concentrated to give 2.9 g of the residue. MeOH (20 mL) and conc. HCl (5 mL) were added and concentrated to afford crude material as a white solid. Recrystallization from MeOH-Et₂O yielded the product [(3-bromo-5-methylphenyl)methyl]amine (3.1 g, 73%) as colorless fine needles. ¹H NMR (400 MHz, D₂O): δ 7.36 (s, 1 H), 7.29 (s, 1 H), 7.10 (s, 1 H), 3.98 (s, 2 H), 2.20 (s, 3 H); ¹³C NMR (400 MHz, D₂O): δ 141.9, 134.6, 132.7, 128.7, 128.5, 122.2, 42.6, 20.4.

Example 25 5-Bromo-2-methylbenzonitrile

Keep the solution of 5-amino-2-methylbenzonitrile (2.0 g, 15.1 mmol) dissolved in water (24 mL) and HBr (74%, 14.4 mL) stirring for 20 min, then the mixture was cooled to 0˜5° C., and the solution of NaNO₂ (1.2 g, 17.4 mmol) in water (13.5 mL) was added. The reaction mixture was stirred for 10 min at 0˜5° C., then was warmed to 40° C. A solution of CuBr (6.5 g, 45.1 mmol) in water (36 mL) and HBr (7.2 mL) was added to the mixture, and it was stirred for 2 h. The mixture was extracted with AcOEt, and the organic layer was washed with saturated NaHCO₃ solution, brine, and dried over Na₂SO₄. The crude product was purified by flask chromatograph (PE:EA=50:1), obtaining 2.3 g of 5-bromo-2-methylbenzonitrile as a white solid (Yield: 77%). ¹H NMR (400 MHz, CDCl₃): δ 7.72 (s, 1 H), 7.59 (d, J=8.0 Hz, 1 H), 7.19 (d, J=8.0 Hz, 1 H), 2.51 (s, 3 H).

Example 26 1,1-Dimethylethyl [(5-bromo-2-methylphenyl)methyl]carbamate

NaBH₄ (2.4 g, 64.3 mmol) was added cautiously to a solution of NiCl₂ (2.8 g, 21.6 mmol), Boc₂O (9.6 g, 44.0 mmol) and 5-bromo-2-methylbenzonitrile (4.2 g, 21.4 mmol) in EtOH (150 mL) at 0° C. within 0.5 h, then stirring for another 40 min after the end of addition. After the reaction had subsided, the mixture was left to stir at room temperature for 0.5 h. Then the solvent was removed and the residue was dissolved in AcOEt and saturated solution of NaHCO₃, then filtered and washed with AcOEt. The combined organic layers were washed with brine and dried over Na₂SO₄. The crude product was purified by flask chromatograph (PE:EA=30:1), obtaining 2.7 g of the product, 1,1-dimethylethyl [(5-bromo-2-methylphenyl)methyl]carbamate, as a white solid (Yield: 42%). ¹H NMR (400 MHz, CDCl₃): δ 7.36 (s, 1 H), 7.28-7.30 (m, 1 H), 7.01 (d, J=16.8 Hz, 1 H), 4.72 (s, 1 H), 4.26-4.30 (m, 2 H), 2.25 (s, 3 H), 1.46 (s, 9 H); ¹³C NMR (100 MHz, CDCl₃): δ 155.9, 139.1, 132.2, 130.6, 127.7, 126.3, 119.9, 42.4, 20.6, 18.7; HPLC: retention time: 4.671 min; purity: 97.2%.

Example 27 1,3-Dibromo-2-methyl-5-nitrobenzene

To a CHCl₃ solution (120 mL) of 1-methyl-4-nitrobenzene (30.0 g, 218.8 mmol), iron powder (3.6 g, 64.5 mmol) was added with mechanically stirring. Then bromine (124.8 g, 40 mL, 780.9 mmol) was added slowly while raising the temperature to 40° C. The mixture was heated to reflux for 48 h. After cooling, the solution was washed with saturated Na₂SO₃ solution, saturated Na₂CO₃ solution, brine, and dried over anhydrous Na₂SO₄. After the solvent was removed, the residual was recrystallized with MeOH, 26.5 g to give yellow crystals of the title compound. Additional 12.3 g of the title compound was obtained by silica column chromatography. Total yield: 60%. ¹H NMR (400 MHz, CDCl₃): δ 2.67 (s, 3 H), 8.38 (s, 2 H).

Example 28 (3,5-Dibromo-4-methylphenyl)amine

1,3-Dibromo-2-methyl-5-nitrobenzene (11.3 g, 38.3 mmol) was dissolved in THF/EtOH (100 mL/100 mL) and then SnCl₂.2H₂O (43.2 g, 191.6 mmol) was added. The mixture was stirred at room temperature for 3 h. After the solvent was removed, NaOH solution (25 g/200 mL) was added, and the mixture was stirred for 1.5 h. The solution was extracted with EtOAc (200 mL×2) and the organic extract dried over anhydrous Na₂SO₄. After removing EtOAc, CH₂Cl₂ was added, and then concentrated HCl (7 mL) was added to form hydrochloric acid salt, which was collected by filtering. The solid was used in subsequent reactions without purification. ¹H NMR (400 MHz, D₂O): δ 2.43 (s, 3 H), 3.61 (br, 2 H), 6.86 (s, 2 H).

Example 29 1,3-Dibromo-2-methylbenzene

A solution of (3,5-dibromo-4-methylphenyl)amine dissolved in water (80 mL) and concentrated HCl (7.5 mL) stirring for 20 min, then the mixture was cooled to 0˜5° C., and the solution of NaNO₂ (3.4 g/30 mL H₂O) was added. The reaction mixture was stirred for 2 h at 0˜5° C., then the suspension was added to hypophosphorous acid solution (50%, 27.9 g), which was cooled to 0° C. The mixture was stirred at room temperature overnight. Then it was extracted with CH₂Cl₂ (100 mL×2) and the combined organic layers were washed with brine (30 mL) and dried over Na₂SO₄. After silica column chromatography, (eluted with petroleum ether), 3.57 g product was obtained, as colorless liquid. ¹H NMR (400 MHz, CDCl₃): δ 2.57 (s, 3 H), 6.89 (t, J=8.0 Hz, 1 H), 7.50 (d, J=8.0 Hz, 2 H).

Example 30 3-Bromo-2-methylbenzoic Acid

To a solution of 1,3-dibromo-2-methylbenzene (6.57 g) in dry THF (100 mL), t-BuLi solution (1.5 M in pentane, 17 mL) was added dropwise at −80° C. After addition, the reaction mixture was stirred between −76˜−78° C. for 2 h. After the mixture was cooled to below −80° C., dry ice was added, and then warmed to room temperature naturally. Solvent was removed, 5% NaOH solution (40 mL) was added, washed with CH₂Cl₂ (10 mL×2). Then the aqueous layer was acidified with concentrated HCl to pH=1, extracted with EtOAc (100 mL×2). The combined organic phase was dried over anhydrous Na₂SO₄. After removing the solvent, the residue was purified by silica column chromatography, (eluted with petrol ether:EtOAc=8:1 to 1:1), to obtain 3.58 g of the product. Yield: 63.4%. ¹H NMR (400 MHz, CDCl₃): δ 2.73 (s, 3 H), 7.15 (t, J=8.0 Hz, 1 H), 7.77 (dd, J=8.0 Hz, J=1.2 Hz, 1 H), 7.94 (dd, J=8.0 Hz, J=1.2 Hz, 1 H).

Example 31 3-Bromo-2-methylbenzamide

3-Bromo-2-methylbenzoic acid (3.7 g) was suspended in dry toluene (50 mL), thionyl chloride (3.8 mL) was added, and then the mixture was heated to reflux for 2 h. After cooling to room temperature, the solvent was removed under reduced pressure. The residue was dissolved in dry THF (10 mL) and toluene (10 mL), added to concentrated ammonia solution (20 mL) and stirred for 1 h. A which solid formed and was filtered off and washed with petrol ether and dried under vacuum to give 1.2 g of product. The filtrate was concentrated to half volume, and then extracted with EtOAc, which was dried over anhydrous Na₂SO₄. After solvent was removed, the resulting white solid was stirred with 20 mL of petroleum ether:2 mL ethyl acetate overnight and filtered to give an additional 1.5 g of product. Product which was used in next step without purification. Yield: 84%. ¹H NMR (400 MHz, CDCl₃): δ 2.52 (s, 3 H), 5.75 (br, 1 H), 5.94 (br, 1 H), 7.08 (t, J=7.6 Hz, 1 H), 7.35 (dd, J=7.4 Hz, J=1.0 Hz, 1 H), 7.62 (dd, J=8.2 Hz, J=1.4 Hz, 1 H).

Example 32 [(3-Bromo-2-methylphenyl)methyl]amine

3-Bromo-2-methylbenzamide (1.4 g) was dissolved with dry THF (15 mL) under nitrogen, then Me₂S.BH₃ (94%, 1.34 mL) was added slowly. After being stirred at room temperature for 1 h, the mixture was heated to 50° C. overnight. When the 3-bromo-2-methylbenzamide disappeared, methanol was added dropwise until there no more air bubble formed. 10 min later, 10% HCl was added dropwise, stirred for 1 h, and then solvent was removed. The white residue was recrystallized with iPrOH to obtain 1.1 g of the product. Yield: 35%. ¹H NMR (400 MHz, DMSO-d6): δ 2.42 (s, 3 H), 4.09 (s, 2 H), 7.20 (t, J=7.8 Hz, 1 H), 7.44 (d, J₁=8.0 Hz, 1 H), 7.63 (d, J₁=7.6 Hz, 1 H), 8.49 (br, 3 H); ¹³C NMR (100 MHz, DMSO-d6): δ 18.8, 19.0, 46.1, 125.2, 127.4, 127.5, 129.0, 129.1, 132.1, 132.5, 134.7, 135.9, 136.1, 136.5; HPLC: retention time: 4.696 min; purity: 96.0%.

Example 33 2-Amino-5-bromo-3-(methyloxy)benzoic Acid

To a solution of 2-amino-3-(methyloxy)benzoic acid (15.0 g, 89.7 mmol) in MeOH (100 mL), NBS (16.8 g, 94.2 mmol) was added at −5° C. The reaction was kept stirring at 0° C. overnight, then put into the ice water under the condition of stirring. A precipitate formed and was filtered over Celite and dried in vacuo to afford 2-amino-5-bromo-3-(methyloxy)benzoic acid (22.0 g, 99%). ¹H NMR (400 MHz, CDCl₃): δ 7.65 (s, 1 H), 6.93 (s, 1 H), 3.87 (s, 3 H).

Example 34 3-Bromo-5-(methyloxy)benzoic Acid

To a solution of 2-amino-5-bromo-3-(methyloxy)benzoic acid (16.40 g, 66.65 mmol) in H₂O (80 mL), conc. HCl (30 mL) and THF (5 mL) were added at 0° C. The reaction was stirred for 30 min, and then NaNO₂ (14.00 g, 202.91 mmol) was cautiously added to the solution. After the reaction mixture had stirred for 2 h, H₃PO₂ (22.00 g, 333.35 mmol) was cautiously added to the solution. The reaction was kept stirring overnight at the room temperature (monitored by TLC), and filtered and rinsed with water (50 mL×2). The formed precipitate was dried to afford 3-bromo-5-(methyloxy)benzoic acid (9.60 g, 62%). ¹H NMR (400 MHz, CDCl₃): δ 7.46 (t, J=1.6 Hz, 1 H), 7.31 (q, J=16.8 Hz, 1 H), 7.21 (t, J=16.8 Hz, 1 H), 3.84 (s, 1 H).

Example 35 3-Bromo-5-(methyloxy)benzamide

To a suspension of 3-bromo-5-(methyloxy)benzoic acid (9.6 g, 41.6 mmol) in toluene (60 mL), thionyl chloride (9.89 g, 83.1 mmol) and DMF (0.10 mL) were added and stirred at 50° C. for 4 h. The reaction was allowed to cooled to room temperature, and then the excess thionyl chloride was removed in vacuo. The residue was dissolved in toluene (50 mL) and the mixture was added to a solution of ammonia (25%, 50 mL). A precipitate formed and was filtered off using Celite, and dried, to afford 3-bromo-5-(methyloxy)benzamide (8.70 g, 90%).

Example 36 {[3-Bromo-5-(methyloxy)phenyl]methyl}amine

To a solution of 3-bromo-5-(methyloxy)benzamide (4.00 g, 17.4 mmol) in THF (60 mL), BH₃.Me₂S (2.64 g, 34.8 mol) was added at 0° C. After the end of the addition, the reaction mixture was kept refluxing overnight (monitored by TLC). After cooling to room temperature, EtOH was cautiously added to the reaction mixture. When there no more air bubbles appeared, the mixture was acidified with 1N HCl to pH=2. Then the mixture was stirred at 50° C. for 4 h, filtered and the filter rinsed with water (20 mL×2). The filtrate was washed with EtOAc (50 mL×3). After addition of 2N NaOH (pH=10), the filtrate was extracted with EtOAc (100 mL×3). The combined extracts were washed with brine, dried over anhydrous sodium sulfate and concentrated to give a mixture of solid and oil. MeOH (10 mL) and conc. HCl (10 mL) were added and concentrated to afford crude material as a white solid. Recrystallization from MeOH-Et₂O gave the captioned product (3.10 g, 71%), {[3-bromo-5-(methyloxy)phenyl]methyl}amine, as colorless fine needles. ¹H NMR (400 MHz, D₂O): δ 7.12-7.09 (m, 2 H), 6.86 (s, 1 H), 3.98 (s, 2 H), 3.69 (s, 3 H); ¹³C NMR (100 MHz, D₂O): δ160.2, 135.8, 124.2, 123.0, 117.8, 114.0, 55.8, 42.5; HPLC: retention time: 5.452 min; purity: 97.4% (HPLC).

Example 37 5-Bromo-2-(methyloxy)benzonitrile

Bromine (13.7 g, 86.0 mmol) in CHCl₃ (20 mL) was added to the solution of 2-(methyloxy)benzonitrile (10.9 g, 81.9 mmol) in CHCl₃ (50 mL). The mixture was refluxed for 29 h. The reaction was allowed to cool to room temperature, and washed with saturated sodium bisulfite (50 mL), and brine (50 mL). The organic layer was dried over anhydrous sodium sulfate. Evaporation of the solvent afforded 5-bromo-2-(methyloxy)benzonitrile (12.4 g, 71%).

Example 38 1,1-Dimethylethyl {[5-bromo-2-(methyloxy)phenyl]methyl}carbamate

NaBH₄ (2.9 g, 75.5 mmol) was cautiously added in several portions to a solution of NiCl₂ (2.6 g, 19.8 mmol), Boc₂O (8.2 g, 37.7 mmol) and 5-bromo-2-(methyloxy)benzonitrile (4.0 g, 18.9 mmol) in dry EtOH (70 mL) at 0° C. Once the reaction had subsided, the mixture was left to stir at room temperature for 3 h. Ethanol was removed under reduced pressure and the residue was dissolved in EtOAc and saturated solution of NaHCO₃, then filtered and repeatedly washed with EtOAc. The combined organic phases were dried using Na₂SO₄. The crude product was purified by flash column chromatography (PE/EA=20:1) to give 1.6 g of the product, 1,1-dimethylethyl {[5-bromo-2-(methyloxy)phenyl]methyl}-carbamate. (Yield: 27%). ¹H NMR (400 MHz, CDCl₃): δ 7.36-7.33 (m, 2 H), 6.74 (d, J=8.8 Hz, 1 H), 4.97 (br, 1 H), 4.27 (d, J=4.8 Hz, 1 H), 3.82 (s, 3 H), 1.45 (s, 9 H); ¹³C NMR (400 MHz, CDCl₃): δ 156.5, 155.8, 131.7, 131.1, 129.3, 111.8, 79.5, 55.5, 39.9, 26.4. HPLC: retention time: 6.592 min; purity: 97.9%.

Example 39 2-Bromo-6-methylphenol

To a solution of o-cresol (20.0 g, 0.19 mol) and iPr₂NH (2.63 mL, 18.5 mmol) in CH₂Cl₂ (500 mL), a solution of NBS (32.9 g, 0.19 mol) in CH₂Cl₂ (500 mL) was added dropwise over 7 h and the mixture was stirred over night at room temperature. The reaction mixture was acidified to pH=1 with conc. sulfuric acid and water (400 mL). The organic layer was separated, dried with Na₂SO₄, and concentrated under reduced pressure. 34.6 g of the crude product was obtained (yield: 97%). ¹H NMR (400 MHz, CDCl₃) δ 2.30 (s, 3 H), 5.54 (s, 2 H), 6.71 (t, J=7.6 Hz, 1 H), 7.05 (d, J=7.6 Hz, 1 H), 7.28 (d, J=8.0 Hz, 1 H).

Example 40 1-Bromo-3-methyl-2-(methyloxy)benzene

To a solution of 2-bromo-6-methylphenol (34.6 g, 0.18 mol) in THF (300 mL), NaH (9.6 g, 0.24 mol, 60%) was added in several portions. After the mixture was stirred for 1 h, Me₂SO₄ (28.0 g, 0.22 mol) was added dropwise, then the mixture was stirred over night. H₂O (50 mL) was added, and the solvent was removed under reduced pressure. Then the residue was dissolved in Et₂O (250 mL), and was washed with NaOH (5%, 100 mL), brine (100 mL), and dried with Na₂SO₄. After removing the solvent, 35.3 g of the crude product was obtained. (Yield: 95%). ¹H NMR (400 MHz, CDCl₃) δ 2.33 (s, 3 H), 3.81 (s, 3 H), 6.88 (t, J=8.0 Hz, 1H), 7.10 (d, J=8.0 Hz, 1 H), 7.30 (d, J=7.6 Hz, 1 H).

Example 41 1-bromo-3-(bromomethyl)-2-(methyloxy)benzene

1-Bromo-3-methyl-2-(methyloxy)benzene (30.3 g, 0.15 mol), NBS (28.2 g, 0.16 mol), and BPO (1.83 g, 7.55 mmol) were suspended in 300 mL of CCl₄, and the mixture was heated to 80° C. over night. After cooled to room temperature, the solution was filtrated and the solid was washed with CCl₄ (30 mL×2). The filtrate was washed with NaHO₃ (aq. 250 mL×2), Na₂CO₃ (aq. 100 mL×2), brine (100 mL) and dried over Na₂SO₄. After removing the solvent, 41.4 g of the crude product was obtained. (Yield: 97.9%).

Example 42 2-{[3-Bromo-2-(methyloxy)phenyl]methyl}-1H-isoindole-1,3(2H)-dione

To a solution of 1-bromo-3-(bromomethyl)-2-(methyloxy)benzene (41.4 g, 0.15 mol) in DMF (350 mL), Potassium Phthalamide (PhtK) (28.8 g, 0.16 mol) was added, and the mixture was heated to 90° C. over night. Then the solvent was removed under reduced pressure. The residue was dissolved in CHCl₃ (300 mL), and filtered. The filtrate was washed with H₂O (100 mL×2), brine (100 mL), and dried over Na₂SO₄. After removing the solvent, the residue was recrystallized with EtOH (200 mL). 26.7 g of the product was obtained as white solid. Yield: 52.1%. ¹H NMR (400 MHz, CDCl₃) δ 3.98 (s, 3 H), 4.95 (s, 2H), 6.93 (t, J=8.0 Hz, 1 H), 7.20 (d, J=0.8 Hz, 1 H), 7.45 (d, J=8.0 Hz, 1 H), 7.72-7.87 (m, 4 H).

Example 43 {[3-Bromo-2-(methyloxy)phenyl]methyl}amine

To a suspension of 2-{[3-bromo-2-(methyloxy)phenyl]methyl}-1H-isoindole-1,3(2H)-dione (26.7 g, 77.2 mmol) in EtOH (300 mL), hydrazine hydrate (7.8 g, 154 mmol) was added, and the reaction mixture was heated to 90° C. for 4 h. After cooled to room temperature, the mixture was filtered and the solid was washed with EtOAc (300 mL×2). The filtrate was concentrated to about 50 mL and filtered again. After removing the solvent, the residue was dissolved in 20 mL of MeOH, and then 1N HCl was added to obtain a white solid. Then the white solid was recrystallized with MeOH-Et₂O to obtain 9.0 g of the product. Yield: 46.3%. ¹H NMR (400 MHz, D₂O) δ 3.79 (s, 3H), 4.13 (s, 2H), 7.02 (t, J=7.6 Hz, 1 H), 7.27 (d, J=8.0 Hz, 1 H), 7.57 (d, J=8.0 Hz, 1 H); ¹³C NMR (100 MHz, D₂O) δ 37.7, 60.2, 115.6, 125.3, 126.6, 128.8, 133.8, 153.7; MS: m/z 254.1 (M⁺); HPLC: retention time: 7.618 min; purity: 98.8%.

Example 44 1-Bromo-3-(bromomethyl)-5-methylbenzene

A mixture of 1-bromo-3,5-dimethylbenzene (25.0 g, 135 mmol), NBS (24.0 g, 135 mmol) and BPO (1.30 g) in CCl₄ (250 mL) was refluxed for 6 h. After cooled to room temperature, the mixture was filtered, and the filtrate was washed successively with saturated sodium bicarbonate (100 mL), water (2×50 mL) and brine (2×50 mL). The combined organic phases were dried (Na₂SO₄) and concentrated in vacuum to give 40.0 g of crude product 1-bromo-3-(bromomethyl)-5-methylbenzene.

Example 45 (3-Bromo-5-methylphenyl)methanol

A mixture of 1-bromo-3-(bromomethyl)-5-methylbenzene (40.0 g, 151 mmol), 1,4-dioxane (150 mL), water (150 mL) and calcium carbonate (37.9 g, 379 mmol) was heated for 16 h at reflux. The mixture was filtered and the filtrate was concentrated in vacuum, then diluted with CH₂Cl₂ (150 mL). The organic layer was washed with HCl (2N, 50 mL) and a solution of saturated sodium bicarbonate (50 mL), dried (Na₂SO₄) and concentrated in vacuum to give 25.0 g of crude product (3-bromo-5-methylphenyl)methanol.

Example 46 3-Bromo-5-methylbenzoic Acid

A solution of KMnO₄ (39.3 g, 249 mmol) in water (600 mL) was added slowly to a solution of (3-bromo-5-methylphenyl)methanol (25.0 g, 124 mmol) in acetone (500 mL), then the mixture was kept at reflux for 60 min. After cooled to room temperature, the mixture was acidified with HCl (2N, 100 mL). The brown precipitate was dissolved by adding a solution of saturated sodium bicarbonate (100 mL), and then acetone was concentrated in vacuum. Ammonia (150 mL) was added. The mixture was filtered over Celite and acidified with concentrated HCl. The product was extracted with diethyl ether (3×150 mL), the combined organic phases were dried (Na₂SO₄) and concentrated in vacuum to obtain 16.0 g of the acid 3-bromo-5-methylbenzoic acid as white crystals (yield: 60%). ¹H NMR (400 MHz, CDCl₃): δ 8.05 (s, 1 H), 7.85-7.84 (m, 1 H), 7.58 (s, 1 H), 2.40 (s, 3 H).

Example 47 3-Bromo-5-methylbenzamide

CDI (42.2 g, 260.4 mmol) was cautiously added to a solution of 3-bromo-5-methylbenzoic acid (16.0 g, 74.4 mmol) in EA (300 mL), and then the mixture was kept at reflux for 3 h. After cooled to room temperature, NH₃ (g) was passed though for 1 h. The mixture was filtered and the organic layer was washed with HCl (10%, 100 mL) and water (100 mL). The organic phase was dried over Na₂SO₄ and concentrated in vacuum to obtain 15.0 g of 3-bromo-5-methylbenzamide as white crystals (yield: 94%). ¹H NMR (400 MHz, CDCl₃): δ 7.73-7.72 (m, 1 H), 7.56-7.55 (m, 1H), 7.50-7.49 (m 1 H), 2.39 (s, 3 H).

Example 48 3-Bromo-5-methylbenzonitrile

To a suspension of 3-bromo-5-methylbenzamide (15.0 g, 70.1 mmol) in CHCl₃ was added phosphorous pentoxide (29.8 g, 210.2 mmol) and the mixture was kept refluxing for 2 days (monitored by TLC). The reaction was allowed to cool to room temperature, and put into the ice water under the condition of stirring. The organic layer was separated and the aqueous layer was extracted with dichloromethane (150 mL×2). The combined extracts were washed with brine, dried over NaSO₄. The product, 3-bromo-5-methylbenzonitrile (7.20 g, 52%), was purified by flash column chromatography. ¹H NMR (400 MHz, CDCl₃): δ 7.60-7.56 (m, 2 H), 7.40-7.39 (m, 1 H), 2.39 (s, 3 H).

Example 49 3-Bromo-5-(bromomethyl)benzonitrile

A mixture of 3-bromo-5-methylbenzonitrile (9.80 g, 45.0 mmol), NBS (8.90 g, 45.0 mmol) and BPO (0.40 g) in CCl₄ (250 mL) was heated for 10 h at reflux. The reaction mixture was cooled to room temperature and filtered, and the organic phase was washed successively with saturated sodium bicarbonate (100 mL), water (2×50 mL) and brine (2×50 mL). The combined organic phases were dried over Na₂SO₄ and concentrated in vacuum to give 12.5 g of crude 3-bromo-5-(bromomethyl)benzonitrile.

Example 50 3-Bromo-5-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]benzonitrile

A suspension of 3-bromo-5-(bromomethyl)benzonitrile (12.5 g, 45.5 mmol), potassium phthalate (7.16 g, 38.6 mmol), and in DMF (100 mL) was stirred under reflux for 4 h. After cooled to room temperature, the solvent was remove under reduced pressure and the residue was dissolved in CHCl₃ (200 mL). The organic layer was washed with water (50 mL×2), dried over Na₂SO₄, and concentrated in vacuum to obtain 15.2 g of the crude product. The product, 3-bromo-5-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]benzonitrile (3.50 g, 23%) was purified by flash column chromatography. ¹H NMR (400 MHz, CDCl₃): δ 7.90-7.87 (m, 2 H), 7.81-7.80 (m, 1 H), 7.77-7.75 (m, 2 H), 7.71-7.70 (m, 1 H), 7.66-7.65 (m, 1 H), 4.83 (s, 2 H).

Example 51 3-(Aminomethyl)-5-bromobenzonitrile

To a suspension of 3-bromo-5-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]benzonitrile (3.50 g, 10.3 mmol) in EtOH (60 mL) was added N₂H₄H₂O (85%, 1.31 g). The mixture was refluxed for 3 h. At room temperature 2 N HCl (20 mL) was added (pH=3), and the mixture was filtered and rinsed with water (20 mL×2). The filtrate was concentrated to about 50 mL and filtered again. After addition of NaHCO₃ (pH=9), the filtrate was extracted with CH₂Cl₂ (50 mL×3). The combined extracts were washed with brine, dried over anhydrous sodium sulfate, and concentrated to give a mixture of colorless solid and colorless oil. 2N HCl in MeOH (50 mL) was added and concentrated to afford crude material as a white solid. Recrystallization from MeOH-Et₂O yielded the product 3-(aminomethyl)-5-bromobenzonitrile (1.40 g, 55%) as colorless fine needles. ¹H NMR (400 MHz, D₂O): δ 7.92 (m, 1 H), 7.84 (m, 1 H), 7.69 (m, 1 H), 4.11 (s, 2 H); ¹³C NMR (400 MHz, D₂O): δ 137.1, 135.9, 135.8, 131.7, 123.0, 117.7, 113.8, 42.0; MS: m/z 209.0 (M⁺−HCl); HPLC: retention time: 9.313 min; purity: 98.4%.

Example 52 (3-Bromophenyl)methanol

To a solution of 3-bromobenzaldehyde (114.8 g, 620.4 mmol) in EtOH (650 mL), NaBH₄ (7.1 g, 186.1 mmol) was added in several portions at 25° C. Then the mixture was stirred for 1 h at room temperature. The reaction was quenched with water (200 mL). After removing EtOH, the residue was dissolved in AcOEt (500 mL), and filtered. The filter was washed with water (150 mL), brine (150 mL), and dried over Na₂SO₄. After removing the solvent, 115.8 g of the title compound was obtained (Yield: 99.8%).

Example 53 {[(3-Bromophenyl)methyl]oxy}(1,1-dimethylethyl)dimethylsilane

TBSCl (18.7 g, 124.3 mmol), Et₃N (14.08 g, 139.2 mmol) and DMAP (194.3 mg, 8.9 mmol) were dissolved in CH₂Cl₂ (120 mL), then the solution was cooled to 0-5° C. (3-bromophenyl)methanol (18.5 g, 99.4 mmol) was added dropwise to the solution. After the addition, the mixture was warmed to room temperature, and stirred for 2 h. 5% HCl was added to the reaction mixture to pH=4-5, then the organic phase was separated, and the aqueous layer was extracted with CH₂Cl₂ (50 mL×2). The combined organic phase was washed with water, and dried over Na₂SO₄. After removing the solvent, 28.5 g of {[(3-Bromophenyl)methyl]oxy}(1,1-dimethylethyl)dimethylsilane was obtained (Yield: 95.1%).

Example 54 [3-({[(1,1-Dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic Acid

To the solution of {[(3-bromophenyl)methyl]oxy}(dimethyl)silane-2,2-dimethylpropane (1:1) (100.0 g, 331.9 mmol) in THF (500 mL), which was cooled to −78° C., n-BuLi (132.7 mL, 331.9 mmol) was added dropwise. The mixture was stirred for 1 h at −78° C., then B(OBu)₃ (107.5 mL, 398.2 mmol) was added one portion. The reaction mixture was warmed to room temperature, and stirred over night. After cooled to 0° C., 5% H₃PO₄ was added to pH=4-5, the mixture was stirred 0.5 h, then was filtered. After removing THF, the residue was extracted with Et₂O (200 mL×2), and the organic layer was dried over Na₂SO₄. After removing the solvent, the residue was added to water, and white solid was precipitated which was dried in vacuo. 65.7 g of [3-({[(1,1-dimethylethyl)-(dimethyl)silyl]oxy}methyl)phenyl]boronic acid was obtained (Yield: 74.5%). ¹H NMR (400 MHz, CDCl₃): δ 0.14 (s, 6 H), 0.98 (s 9 H), 4.88 (s, 2 H), 7.49-7.59 (m, 2 H), 8.14 (d, J=7.6 Hz, 1 H), 8.19 (s, 1 H).

Example 55 1,1-Dimethylethyl [(3-bromo-4-cyanophenyl)methyl]carbamate

To a suspension of 4-(aminomethyl)-2-bromobenzonitrile (2.2 g, 8.9 mmol) and Na₂CO₃ (2.4 g, 21.4 mmol) in CH₂Cl₂ (50 mL), the solution of Boc₂O (2.1 g, 9.8 mmol) in CH₂Cl₂ (10 mL) was added dropwise. Then the reaction mixture was stirred overnight at room temperature. After filtration, the solid was washed with CH₂Cl₂ (20 mL×2), and then the filtrate was washed with water (20 mL×2), brine (20 mL×2) and dried over Na₂SO₄. After removing the solvent, 2.6 g of 1,1-dimethylethyl [(3-bromo-4-cyanophenyl)methyl]carbamate was obtained (yield: 94%).

Example 56 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(methyl)silyl]oxy}methyl)-6-methyl-3-biphenylyl]methyl}carbamate-ethane (1:1)

Pd(OAc)₂ (56.3 mg, 0.25 mmol), PPh₃ (263.0 mg, 1.0 mmol), K₂CO₃ (1.7 g, 12.5 mmol) and 1,1-dimethylethyl [(3-bromo-4-cyanophenyl)methyl]carbamate (2.6 g, 8.4 mmol) were suspended in 1,4-dioxane (30 mL). After the mixture was heated to 80° C. for 15 min, [3-(hydroxymethyl)phenyl]boronic acid-(1,1-dimethylethyl)(trimethyl)silane (2.7 g, 10.0 mmol) was added. Then the reaction mixture was stirred over night at 100° C. After cooled to room temperature, the solvent was removed under reduced pressure. The residue was dissolved in CH₂Cl₂ (50 mL), then washed with water (20 mL), brine (20 mL), and dried over Na₂SO₄. After the solvent was removed, the crude product was purified on Al₂O₃ column chromatography, eluting with CH₂Cl₂. 2.6 g of the product, 1,1-dimethylethyl{[3′-({[(1,1-dimethylethyl)(methyl)silyl]oxy}methyl)-6-methyl-3-biphenylyl]methyl}carbamate-ethane, was obtained (yield: 60%). ¹H NMR (400 MHz, CDCl₃): δ 0.12 (s, 6 H), 0.95 (s 9 H), 1.46 (s, 9 H), 4.41-4.42 (m, 2 H), 4.81 (s, 2 H), 7.37-7.47 (m, 6 H), 7.71 (d, J=8.0 Hz, 1 H); ¹³C NMR (100 MHz, CDCl₃): δ −5.3, 18.4, 25.9, 28.3, 44.2, 64.7, 80.0, 109.9, 118.6, 126.1, 126.2, 126.4, 127.3, 128.6, 134.0, 137.9, 142.0, 144.5, 145.8, 155.8; HPLC: retention time: 9.500 min; purity: 95.2% (HPLC).

Example 57 3′-({[(1,1-Dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-fluoro-3-biphenylcarbonitrile

3-Bromo-5-fluorobenzonitrile (5.00 g, 25.0 mmol), Pd(OAc)₂ (0.15 g), PPh₃ (0.60 g) and K₂CO₃ (5.18 g, 37.5 mmol) were dissolved in dioxane (60 mL). The mixture was heated at 70° C. for 30 min, then [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (7.99 g, 30.0 mmol) was added. The mixture reaction was stirred at reflux overnight. The solvent was removed under reduced pressure, then diluted with CH₂Cl₂ (100 mL). The organic layer was washed with water (50 mL) and brine (50 mL). And the organic layer was dried over Na₂SO₄. The product 3′-({[(1,1-dimethylethyl) (dimethyl)silyl]oxy}methyl)-5-fluoro-3-biphenylcarbonitrile (5.10 g, 60%) was purified by flash column chromatography. ¹H NMR (400 MHz, CDCl₃): δ 7.67 (t, J=5.2 Hz, 1 H), 7.54-7.32 (m, 5 H), 4.81 (s, 2 H), 0.94 (s, 9 H), 0.13 (s, 6 H).

Example 58 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy methyl)-5-fluoro-3-biphenylyl]methyl}carbamate

NaBH₄ (3.57 g, 94.3 mmol) was cautiously added to a solution of NiCl₂ (1.83 g, 14.1 mmol), Boc₂O (6.03 g, 27.6 mmol) and 3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-fluoro-3-biphenylcarbonitrile (4.60 g, 13.5 mmol) in absolute ethanol (70 mL) at 0° C. (vigorous reaction with the formation of a black precipitate). Once the reaction had subsided the mixture was left to stir, at room temperature, for 30 min. Ethanol was removed under reduced pressure and the precipitate dissolved in EtOAc and NaHCO₃, filtered and repeatedly washed with EtOAc. The combined organic phases were dried (Na₂SO₄). The product was purified by flash column chromatography to yield 1,1-dimethylethyl {[3′-({[(1,1-dimethylethyl)-(dimethyl)silyl]oxy}methyl)-5-fluoro-3-biphenylyl]methyl}carbamate (1.90 g, 32%). ¹H NMR (400 MHz, CDCl₃): δ 7.51-7.16 (m, 6 H), 6.97 (d, J=5.2 Hz, 1 H), 4.91 (s, 1 H), 4.80 (s, 2 H), 4.38-4.37 (m, 2 H), 1.53 (s, 9 H), 0.96 (s, 9 H), 0.12 (s, 6 H); ¹³C NMR (400 MHz, CDCl₃): δ 156.0, 143.9, 142.3, 139.8, 129.0, 125.8, 124.9, 121.8, 113.1, 79.9, 65.0, 44.4, 28.5, 26.1, 18.6, −5.1; HPLC: retention time: 4.709 min; purity: 97.9% (HPLC).

Example 59 3′-({[(1,1-Dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-fluoro-3-biphenylcarboxamide

Pd(OAc)₂ (123.6 mg, 0.55 mmol), PPh₃ (557.5 mg, 2.2 mmol), K₂CO₃ (3.8 g, 27.5 mmol) and 3-bromo-2-fluorobenzamide (4.0 g, 18.4 mmol) were suspended in 1,4-dioxane (30 mL). After the mixture was heated to 80° C. for 15 min, [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (5.9 g, 22.0 mmol) was added. Then the reaction mixture was stirred over night at 100° C. After cooled to room temperature, the solvent was removed under reduced pressure. The residue was dissolved in CH₂Cl₂ (50 mL), then was washed with water (20 mL), brine (20 mL), and dried over Na₂SO₄. After the solvent was removed, the crude product was purified on Al₂O₃ column chromatography, eluting with CH₂Cl₂/CH₃OH (300:1). 4.1 g of the product, 3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-fluoro-3-biphenylcarboxamide, was obtained (yield: 63%). ¹H NMR (400 MHz, CDCl₃): δ 0.12 (s, 6 H), 0.95 (s, 9 H), 4.81 (s, 2 H), 7.25-8.12 (m, 7 H).

Example 60 [3′-(Aminomethyl)-2′-fluoro-3-biphenylyl]methanol

To a solution of 3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-fluoro-3-biphenylcarboxamide (3.8 g, 10.6 mmol) in THF (40 mL) which was cooled to 0° C., BH₃Me₂S (14.0 mL, 21.2 mmol) was added dropwise. Then the reaction was stirred at 50° C. overnight. The reaction was quenched by adding HCl (10 mL, 3 N) and the result mixture was stirred for 2 h before THF was removed under vacuum. The aqueous layer was extracted with AcOEt (30 mL) then the pH was adjusted to around 9.0 by adding Na₂CO₃. The aqueous layer was extracted with AcOEt (50 mL×2), and dried over Na₂SO₄. After the solvent was removed, the crude product was purified on Al₂O₃ column chromatography, eluting with CH₂Cl₂/EA (10:1). 0.83 g of the product, [3′-(aminomethyl)-2′-fluoro-3-biphenylyl]methanol, was obtained (yield: 33%). ¹H NMR (400 MHz, DMSO): δ 3.80 (s, 2 H), 4.56 (s, 2 H), 7.23-7.51 (m, 7 H); ¹³C NMR (400 MHz, DMSO): δ 62.8, 124.3, 125.8, 126.9, 127.2, 128.0, 128.1, 128.3, 128.4, 128.6, 131.6, 131.8, 135.6, 142.9, 155.6, 158.1; HPLC: retention time: 4.053 min; purity: 98.6% (HPLC).

Example 61 1,1-Dimethylethyl [(3-bromo-2-fluorophenyl)methyl]carbamate

To a suspension of [(3-bromo-2-fluorophenyl)methyl]amine (5.0 g, 20.3 mmol) and Na₂CO₃ (5.5 g, 51.9 mmol) in CH₂Cl₂ (100 mL), the solution of Boc₂O (4.5 g, 20.6 mmol) in CH₂Cl₂ (10 ml) was added dropwise. Then the reaction mixture was stirred overnight at room temperature. After filtration, the solid was washed with CH₂Cl₂ (50 mL×2), and then the filtrate was washed with water (70 mL×2), brine (70 mL×2) and dried over Na₂SO₄. After removing the solvent, 5.6 g of 1,1-dimethylethyl [(3-bromo-2-fluorophenyl)methyl]carbamate was obtained (yield: 94%).

Example 62 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-fluoro-3-biphenylyl]methyl}carbamate

Pd(OAc)₂ (88.9 mg, 0.39 mmol), PPh₃ (415.0 mg, 1.6 mmol), K₂CO₃ (2.7 g, 19.8 mmol) and 1,1-dimethylethyl [(3-bromo-2-fluorophenyl)methyl]carbamate (4.0 g, 13.2 mmol) were suspended in 1,4-dioxane (50 mL). After the mixture was heated to 80° C. for 15 min, [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (4.2 g, 15.8 mmol) was added. Then the reaction mixture was stirred over night at 100° C. After cooled to room temperature, the solvent was removed under reduced pressure. The residue was dissolved in CH₂Cl₂ (80 mL), then was washed with water (30 mL), brine (30 mL), and dried over Na₂SO₄. After the solvent was removed, the crude product was purified on Al₂O₃ column chromatography, eluting with PE/EA (20:1). 1.98 g of the product, 1,1-dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-fluoro-3-biphenylyl]methyl}carbamate, was obtained (yield: 34%). ¹H NMR (400 MHz, CDCl₃): δ 0.12 (s, 6 H), 0.95 (s, 9 H), 4.42-4.43 (m, 2 H), 4.80 (s, 2 H), 4.96 (s, 1 H), 7.17-7.48 (m, 7 H); ¹³C NMR (400 MHz, CDCl₃): δ −5.3, 18.4, 25.9, 28.3, 38.9, 64.8, 124.2, 125.4, 126.7, 127.6, 128.3, 128.7, 129.7, 129.2, 129.7, 135.4, 141.6, 155.8; HPLC: retention time: 4.630 min; purity: 99.4% (HPLC).

Example 63 2-{[3′-({[(1,1-Dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-methyl-3-biphenylyl]methyl}-1H-isoindole-1,3(2H)-dione

Pd(OAc)₂ (102.0 mg, 0.45 mmol, 0.03 eq.), PPh₃ (476.4 mg, 1.82 mmol, 0.12 eq.), K₂CO₃ (3.14 g, 22.7 mmol, 1.50 eq.) and 2-[(3-bromo-5-methylphenyl)methyl]-1H-isoindole-1,3(2H)-dione (5.00 g, 13.1 mmol, 1.00 eq.) were suspended in anhydrous 1,4-dioxane (30 mL) under nitrogen. After the mixture was heated to 60° C. for 10 min, [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (4.84 g, 18.2 mmol, 1.20 eq.) was added. Then the reaction mixture was stirred over night at 100° C. After cooled to room temperature, the solvent was removed under reduced pressure. Then water (25 mL) was added, extracted with CH₂Cl₂ twice (70 mL, 50 mL). The organic layer was washed with brine (20 mL×2), dried over anhydrous Na₂SO₄. After the solvent was removed, the crude product was purified on silica column chromatography, eluting with PE/EA (20:1 to 10:1), 4.2 g of product 2-{[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-methyl-3-biphenylyl]methyl}-1H-isoindole-1,3(2H)-dione was obtained, as a colorless liquid. Yield: 59%. ¹H NMR (400 MHz, CDCl₃): δ 0.11 (s, 6 H), 0.95 (s, 9 H), 2.38 (s, 3 H), 4.79 (s, 2 H), 4.87 (s, 2 H), 7.23 (s, 1 H), 7.31-7.49 (m, 6 H), 7.70-7.72 (m, 2 H), 7.84-7.86 (m, 2 H).

Example 64 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-methyl-3-biphenylyl]methyl}carbamate

2-{[3′-({[(1,1-Dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-methyl-3-biphenylyl]methyl}-1H-isoindole-1,3(2H)-dione (4.15 g, 8.8 mmol, 1.0 eq.) was dissolved in ethanol (84 mL), then hydrazine hydrate (85%, 1.1 g, 2.0 eq.) was added. The mixture was heated to reflux, after 5.5 h, reaction completed. Filtered to remove 2,3-dihydrophthalazine-1,4-dione, filtration was concentrated. Then the residue was dissolved in THF (50 mL), and filtered. After removing the solvent, 2.5 g of colorless oil was obtained. The oil was dissolved in CH₂Cl₂ (50 mL) and THF (5 mL), followed by adding anhydrous Na₂CO₃ (1.4 g, 13.2 mmol). After stirred for 15 min, the CH₂Cl₂ solution (20 mL) of Boc₂O (2.1 g, 9.6 mmol) was added dropwise. When stirred for 30 min, reaction completed. Filtered, then solvent was removed. The residual was purified by chromatography (petroleum ether:ethyl acetate=30:1) on alumina basic, finally, 1.7 g of the above named product was obtained. Yield was 43.2% overall the two steps. ¹H NMR (400 MHz, CDCl₃): δ 0.12 (s, 6 H), 0.96 (s, 9 H), 1.47 (s, 9 H), 2.40 (s, 3 H), 4.35 (d, J=6.0 Hz, 2 H), 4.80 (s, 1 H), 4.84 (br, 1 H), 7.09 (s, 1 H), 7.29-7.52 (m, 6 H); ¹³C NMR (100 MHz, CDCl₃): δ −5.2, 18.4, 21.4, 25.9, 28.4, 64.9, 79.4, 123.4, 124.8, 125.0, 125.7, 127.0, 127.2, 128.6, 138.7, 139.2, 140.9, 141.6, 141.9, 155.9; HPLC: retention time: 5.296 min; purity: 99.1% (HPLC).

Example 65 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-4-methyl-3-biphenylyl]methyl}carbamate

1,1-Dimethylethyl [(5-bromo-2-methylphenyl)methyl]carbamate (2.7 g, 9.0 mmol), Pd(OAc)₂ (81 mg, 0.36 mmol), dicyclohexyl[2′-(methyloxy)-1,1′-binaphthalen-2-yl]phosphane (216 mg, 0.45 mmol) and K₃PO₄ (2.5 g, 11.7 mmol) were dissolved in dioxane (50 mL). The mixture was heated at 80° C. for 30 min, and then [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (3.1 g, 11.7 mmol) was added. The mixture reaction was stirred at reflux for two days. The solvent was removed under reduced pressure, then diluted with CH₂Cl₂ (100 mL). The organic layer was washed with water (30 mL), brine (30 mL), and dried over Na₂SO₄. After removing the solvent, 3.6 g of the crude product 1,1-dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-4-methyl-3-biphenylyl]methyl}carbamate was obtained. Yield: 90%.

Example 66 1,1-Dimethylethyl {[3′-(hydroxymethyl)-4-methyl-3-biphenylyl]methyl}carbamate

To a solution of 1,1-dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)-silyl]oxy}methyl)-4-methyl-3-biphenylyl]methyl}carbamate (3.60 g, 8.2 mmol) in THF (30 mL), the solution of nBu₄NF (2.34 g, 9.0 mmol) in THF (20 mL) was added, and then the mixture was stirred at room temperature over night. The solvent was removed under reduced pressure, then diluted with CH₂Cl₂ (50 mL). The organic layer was washed with water (15 mL×2), brine (15 mL×2), and dried over Na₂SO₄. The product 1,1-dimethylethyl {[3′-(hydroxymethyl)-4-methyl-3-biphenylyl]methyl}carbamate (1.5 g) was purified by flash column chromatography (PE:EA=4:1). (Yield: 53%) 1H NMR, 13C NMR, HPLC, MS¹H NMR (400 MHz, CDCl₃): δ1.47 (s, 9 H), 2.37 (s, 3 H), 4.37 (d, J=5.6 Hz, 2 H), 4.76 (m, 3 H), 7.23-7.58 (m, 7 H); ¹³C NMR (100 MHz, CDCl₃): δ 18.6, 28.4, 42.9, 65.3, 79.5, 125.6, 125.7, 126.2, 126.8, 129.0, 130.9, 135.5, 138.9, 141.2, 141.4, 155.8; HPLC: retention time: 14.965 min; purity: 95.4%; MS: m/z 327 (M⁺).

Example 67 1,1-Dimethylethyl [(3-bromo-2-methylphenyl)methyl]carbamate

[(3-Bromo-2-methylphenyl)methyl]amine (4.0 g, 17.0 mmol) was suspended in CH₂Cl₂ (50 mL), then sodium carbonate (4.8 g, 45.3 mmol) was added. After stirred for 15 min, the solution of Boc₂O (4.0 g, 18.3 mmol) in CH₂Cl₂ (20 mL) was added, and then the mixture was stirred overnight. After the solvent was removed, the residue was dissolved in CH₂Cl₂ (40 mL). The solution was washed with water (15 mL), brine (15 mL) and dried over anhydrous Na₂SO₄. After silica column chromatography, (eluted with petroleum ether:EtOAc=20:1 to 5:1), 1.3 g of the product, 1,1-dimethylethyl [(3-bromo-2-methylphenyl)methyl]carbamate, was obtained. Yield: 35%. ¹H NMR (400 MHz, CDCl₃): δ 1.45 (s, 9 H), 2.40 (s, 3 H), 4.34 (d, J=6.0 Hz, 2 H), 4.71 (br, 1 H), 7.02 (t, J=8.0 Hz, 1 H), 7.19 (d, J=7.6 Hz, 1 H), 7.48 (dd, J=7.8 Hz, J=0.6 Hz, 1 H).

Example 68 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-methyl-3-biphenylyl]methyl}carbamate

Pd(OAc)₂ (25.8 mg, 0.115 mmol), PPh₃ (120.6 mg, 0.46 mmol), K₂CO₃ (794.0 mg, 5.75 mmol) and 1,1-dimethylethyl [(3-bromo-2-methylphenyl)methyl]carbamate (1.15 g, 3.85 mmol) were suspended in anhydrous 1,4-dioxane (20 mL) under nitrogen. After the mixture was heated to 60° C. for 30 min, [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (2.04 g, 4.62 mmol) was added. Then the mixture was stirred overnight at 100° C. After cooled to room temperature, the solvent was removed under reduced pressure. The residue was dissolved in EtOAc (20 mL), and then the solution was washed with water (7 mL), brine (7 mL), and, dried over anhydrous Na₂SO₄. After silica column chromatography, eluted with PE:EA=30:1, 1.1 g of the product was obtained, as a colorless liquid. (Yield: 65%). ¹H NMR (400 MHz, CDCl₃): 0.10 (s, 6 H), 0.94 (s, 9 H), 1.47 (s, 9 H), 2.19 (s, 3 H), 4.38 (d, J=5.2 Hz, 2 H), 4.76 (br, 1 H), 4.78 (s, 2 H), 7.14-7.39 (m, 7 H). ¹³C NMR (100 MHz, CDCl₃): δ −5.2, 16.1, 18.4, 25.9, 28.4, 43.4, 64.9, 79.4, 124.6, 125.6, 127.0, 127.9, 129.3, 133.7, 136.9, 141.2, 141.9, 143.0, 155.7; HPLC: retention time: 4.987 min; purity: 98.9% (HPLC).

Example 69 1,1-Dimethylethyl {[3-bromo-5-(methyloxy)phenyl]methyl}carbamate

To a solution of {[3-bromo-5-(methyloxy)phenyl]methyl}amine (1.3 g, 6.0 mmol) in CH₂Cl₂ (15 mL), a solution of NaOH (264.7 mg, 6.6 mmol) in H₂O (6 mL) was added, and then the solution of Boc₂O (1.44 g, 6.6 mmol) in CH₂Cl₂ (20 mL) was added dropwise. The reaction mixture was stirred overnight at 3 h. The aqueous layer was extracted with CH₂Cl₂ (10 mL), then the combined organic phase was washed with brine (10 mL×2) and dried over Na₂SO₄. The product 1,1-dimethylethyl {[3-bromo-5-(methyloxy)phenyl]methyl}carbamate (1.4 g) was purified by flash column chromatography (PE:EA=10:1). Yield: 74%.

Example 70 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-(methyloxy)-3-biphenylyl]methyl}carbamate

1,1-Dimethylethyl {[3-bromo-5-(methyloxy)phenyl]methyl}carbamate (1.4 g, 4.43 mmol), Pd(OAc)₂ (70.0 mg, 0.14 mmol), dicyclohexyl[2′-(methyloxy)-1,1′-binaphthalen-2-yl]phosphane (84.0 mg, 0.175 mmol) and K₃PO₄ (1.2 g, 5.31 mmol) were dissolved in dioxane (30 mL). The mixture was heated at 80° C. for 30 min, and then [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (1.5 g, 5.76 mmol) was added. The mixture reaction was stirred at reflux for two days. The solvent was removed under reduced pressure, then diluted with CH₂Cl₂ (100 mL). The organic layer was washed with water (20 mL), brine (20 mL), and dried over Na₂SO₄. The product 1,1-dimethylethyl[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-(methyloxy)-3-biphenylyl]methyl}carbamate (1.5 g) was purified by flash column chromatography (PE:EA=15:1). Yield: 74%.

Example 71 1,1-Dimethylethyl {[3′-(hydroxymethyl)-5-(methyloxy)-3-biphenylyl]methyl}carbamate

To a solution of 1,1-dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-5-(methyloxy)-3-biphenylyl]methyl}carbamate (1.5 g, 3.28 mmol) in THF (20 mL), the solution of nBu₄NF (0.94 g, 3.61 mmol) in THF (10 mL) was added, and then the mixture was stirred at room temperature over night. After the solvent was removed under reduced pressure, the residue was diluted with EtOAc (30 mL). The organic layer was washed with water (10 mL×2), brine (10 mL×2), and dried over Na₂SO₄. The product 1,1-dimethylethyl {[3′-(hydroxymethyl)-5-(methyloxy)-3-biphenylyl]methyl}carbamate (0.9 g) was purified by flash column chromatography (PE:EA=3:1). (Yield: 80%). ¹H NMR (400 MHz, CDCl₃): δ 1.47 (s, 9 H), 3.86 (s, 3 H), 4.35 (d, J=7.6 Hz, 2 H), 4.76 (s, 2 H), 4.89 (s, 1 H), 6.83 (s, 1 H), 7.02 (s, 1 H), 7.09 (s, 1 H), 7.34-7.58 (m, 4H); ¹³C NMR (100 MHz, CDCl₃): δ 28.4, 44.7, 55.4, 65.2, 79.6, 111.9, 118.7, 125.8, 126.1, 126.4, 129.0, 140.9, 141.2, 141.5, 142.8, 156.0, 160.3. HPLC: retention time: 11.558 min; purity: 98.7%; MS: m/z 343 (M⁺).

Example 72 1,1-Dimethylethyl {[3-bromo-2-(methyloxy)phenyl]methyl}carbamate

To a suspension of {[3-bromo-2-(methyloxy)phenyl]methyl}amine (5.0 g, 19.8 mmol) and Na₂CO₃ (5.3 g, 49.5 mmol) in CH₂Cl₂ (100 mL), the solution of Boc₂O (4.8 g, 21.8 mmol) in CH₂Cl₂ (10 mL) was added dropwise. Then the reaction mixture was stirred overnight at room temperature. After filtration, the solid was washed with CH₂Cl₂ (50 mL×2), and then the filtrate was washed with water (70 mL×2), brine (70 mL×2) and dried over Na₂SO₄. After removing the solvent, 5.5 g of 1,1-dimethylethyl {[3-bromo-2-(methyloxy)phenyl]methyl}carbamate was obtained (yield: 87.8%).

Example 73 1,1-Dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-(methyloxy)-3-biphenylyl]methyl}carbamate

1,1-Dimethylethyl {[3-bromo-2-(methyloxy)phenyl]methyl}carbamate (5.2 g, 16.4 mmol), Pd(OAc)₂ (110.4 mg, 0.49 mmol), dicyclohexyl[2′-(methyloxy)-1,1′-binaphthalen-2-yl]phosphane (317.2 mg, 0.66 mmol) and K₃PO₄ (4.2 g, 19.7 mmol) were dissolved in dioxane (60 mL). The mixture was heated at 80° C. for 30 min, and then [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (5.2 g, 19.7 mmol) was added. The mixture reaction was stirred at reflux overnight. The solvent was removed under reduced pressure, then diluted with CH₂Cl₂ (100 mL). The organic layer was washed with water (30 mL), brine (30 mL), and dried over Na₂SO₄. The product 1,1-dimethylethyl {[3′-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-2-(methyloxy)-3-biphenylyl]-methyl}carbamate (5.0 g) was purified by flash column chromatography (PE:EA=15:1). (Yield: 72%). ¹H NMR (400 MHz, CDCl₃): δ 0.11 (s, 6 H), 0.94 (s, 9 H), 1.46 (s, 9 H), 3.37 (s, 3 H), 4.40 (d, J=6 Hz, 2 H), 4.80 (s, 2 H), 5.04 (s, 1 H), 7.12-7.51 (m, 7 H); ¹³C NMR (100 MHz, CDCl₃): δ −5.2, 18.4, 25.9, 28.4, 40.3, 60.4, 65.0, 124.2, 125.0, 126.6, 127.5, 128.3, 128.4, 130.6, 132.3, 134.8, 138.2, 141.5, 155.8; HPLC: retention time: 4.348 min; purity: 99.9%; MS: m/z 453 (M⁺), 344 (M⁺−TBS).

Example 75 1,1-Dimethylethyl [(3-bromo-5-cyanophenyl)methyl]carbamate

To a suspension of 3-(aminomethyl)-5-bromobenzonitrile (1.6 g, 6.5 mmol) and Na₂CO₃ (1.7 g, 16.2 mmol) in CH₂Cl₂ (25 mL), the solution of Boc₂O (1.6 g, 7.1 mmol) in CH₂Cl₂ (10 mL) was added dropwise. Then the reaction mixture was stirred overnight at room temperature. After filtration, the solid was washed with CH₂Cl₂ (10 mL×2), and then the filtrate was washed with water (20 mL×2), brine (20 mL×2) and dried over Na₂SO₄. After removing the solvent, 1.9 g of 1,1-dimethylethyl [(3-bromo-5-cyanophenyl)methyl]carbamate was obtained (yield: 94.5%).

Example 76 1,1-Dimethylethyl {[5-cyano-3′-({[dimethyl(1-methylethyl)silyl]oxy}methyl)-3-biphenylyl]methyl}carbamate-ethane (1:1)

1,1-Dimethylethyl [(3-bromo-5-cyanophenyl)methyl]carbamate (1.90 g, 6.11 mmol), Pd(OAc)₂ (76 mg), PPh₃ (228 mg) and K₂CO₃ (1.27 g, 9.16 mmol) were dissolved in dioxane (50 mL). The mixture was heated at 70° C. for 30 min, and then [3-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)phenyl]boronic acid (2.11 g, 7.94 mmol) was added. The mixture reaction was stirred at reflux overnight. The solvent was removed under reduced pressure, then diluted with CH₂Cl₂ (100 mL). The organic layer was washed with water (30 mL) and brine (30 mL). And the organic layer was dried over Na₂SO₄. The product 1,1-dimethylethyl {[5-cyano-3′-({[dimethyl(1-methylethyl)silyl]oxy}methyl)-3-biphenylyl]methyl}carbamate-ethane (2.0 g) was purified by flash column chromatography. Yield: 72%. ¹H NMR (400 MHz, CDCl₃): δ 0.12 (s, 6 H), 0.96 (s, 9 H), 1.47 (s, 9 H), 4.41 (d, J=6 Hz, 2 H), 4.81 (s, 2 H), 7.38-7.44 (m, 3 H), 7.50 (s, 1 H), 7.55 (s, 1 H), 7.70 (s, 1 H), 7.75 (s, 1H); ¹³C NMR (100 MHz, CDCl₃): δ −5.2, 18.4, 25.9, 28.3, 43.4, 64.7, 80.1, 113.1, 118.7, 124.7, 125.6, 126.1, 129.0, 129.2, 129.6, 130.4, 133.4, 134.8, 138.6, 141.2, 142.5, 142.9, 155.9; HPLC: retention time: 4.670 min; purity: 94.4%; MS: m/z 453 (M⁺, 32), 339 (M⁺−TBS, 100).

Example 77 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

To a solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (0.6 g, 2 mmol) stirring at room temperature under argon in DMF (150 mL) was added 3-[(methyloxy)carbonyl]benzoic acid (0.356 g, 2 mmol), followed by PyBOP (1.14 g, 2.2 mmol), and N,N-diisopropylethylamine (1.38 mL, 8 mmol). The mixture was stirred at room temperature for 17 h. The solvent was pumped off, and the residue taken up in CH₂Cl₂ (300 mL), and washed with water (300 mL). The organic phase dried over anhydrous Na₂SO₄, filtered and concentrated. This was then CombiFlashed on a 40 gram silica column eluted with 20-100% EtOAc to afford the title compound. LC-MS m/z 465.8 (M+H)⁺, 1.49 min (ret time).

Example 78 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (0.466 g, 1 mmol), lithium hydroxide (0.237 g, 10 mmol), THF (30 mL), and water (10 mL) were combined and stirred under argon at room temperature overnight. Solvents were pumped off, and the residue taken up in ethyl acetate and water. The mixture adjusted to pH 2.5 with HCl, and extracted with a large volume of ethyl acetate. Organic phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 451.8 (M+H)⁺, 1.37 min (ret time).

Example 79 [5′-(Aminomethyl)-2′-fluoro-3-biphenylyl]methanol

[(3-Bromo-4-fluorophenyl)methyl]amine hydrochloride (0.795 g, 3.29 mmol), [3-(hydroxymethyl)phenyl]boronic acid (0.5 g, 3.29 mmol), potassium carbonate (2.275 g, 16.5 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.114 g, 0.1 mmol) were combined in dioxane (10 mL) and water (3 mL). The mixture was microwaved at 150° C. for 30 min. The solvents were concentrated and the residue taken up in EtOAc and H₂O. The aqueous phase was extracted 2× with EtOAc. The combined organic phases were dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by CombiFlash on a silica gel column eluted with 0-100% CH₂Cl₂/CH₂Cl₂: 20% MeOH: 1% NH₄OH to afford the title compound as viscous oil. LC-MS m/z 231.8 (M+H)⁺, 1.03 min (ret time).

Example 80 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-amino)carbonyl]benzoic acid (0.045 g, 0.1 mmol) in CH₂Cl₂ (2 mL) was stirred under argon at room temperature. HBTU (0.038 g, 0.1 mmol) was added followed by [5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methanol (0.0231 g, 0.1 mmol) and triethylamine (28 μL, 0.2 mmol). The resulting mixture was stirred for three days. The solvents were pumped off and the residue was purified on the CombiFlash (12 g silica gel column) eluted with 70-100% EtOAc/hexane) to afford the title compound. LC-MS m/z 664.8 (M+H)⁺, 1.75 min (ret time).

Example 81 N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.039 g, 0.059 mmol), thionyl chloride (0.2 mL, 2.7 mmol), and CH₂Cl₂ (2 ml) were combined and warmed while stirring to 40° C. for 2 h, and then allowed to stir at room temperature overnight. The solvent was pumped off to afford the title compound. LC-MS m/z 682.8 (M+H)⁺, 2.14 min (ret time).

Example 82 Methyl 3-[({[6-Fluoro-3′-(hydroxymethyl)-3-biphenylyl]-methyl}amino) carbonyl]benzoate

To 3-[(methyloxy)carbonyl]benzoic acid (0.18 g, 1 mmol) in CH₂Cl₂ (25 mL) was added HBTU (0.379 g, 1 mmol), followed by [5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methanol (0.231 g, 1 mmol), and triethylamine (0.278 mL, 2 mmol). The mixture was stirred under argon overnight. The reaction mixture was then concentrated to dryness, and taken up in EtOAc and H₂O. The organic phase was washed 2× with H₂O, 1× with brine, dried and concentrated. The residue was absorbed on Isolute® Sorbent, and CombiFlashed on a 12 g column eluted with 0-100% EtOAc/hexane to afford the title compound as a white solid. LC-MS m/z 393.8 (M+H)⁺, 1.98 min (ret time); analytical HPLC shows 97.2% purity, (ret time 7.205 min).

Example 83 3-[({[6-Fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-amino)carbonyl]benzoic Acid

Methyl 3-[({[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}amino)-carbonyl]benzoate (0.309 g, 0.786 mmol), lithium hydroxide (0.0282 g, 1.2 mmol), THF (6 mL), and water (2 mL) were combined and stirred under argon at room temperature overnight. Solvents were pumped off, the residue adjusted to pH 2 with 1N HCl, and extracted with EtOAc, washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated to afford the title compound as a white solid. LC-MS m/z 379.8 (M+H)⁺, 1.69 min (ret time); mp 169-171° C.; analytical HPLC shows 93.2% purity, (ret time 7.059 min).

Example 84 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

3-[({[6-Fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}amino)carbonyl]benzoic acid (0.06 g, 0.158 mmol), 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (0.048 g, 0.158 mmol), HBTU (0.06 g, 0.158 mmol), TEA (44 μL, 0.316 mmol) were combined in CH₂Cl₂ (10 mL) and stirred under argon overnight. Aqueous workup followed by CombiFlash purification on a 12 g column eluted with 0-8% MeOH/CH₂Cl₂ afforded the title compound. LC-MS m/z 664.8 (M+H)⁺, 1.80 min (ret time).

Example 85 1,1-Dimethylethyl (1S,4S)-5-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-amino)carbonyl]phenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2,5-diazabicyclo [2.2.1]heptane-2-carboxylate

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.068 g, 0.1 mmol) and 1,1-dimethylethyl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (0.099 g, 0.5 mmol) were combined in THF (1 mL) and microwaved at 150° C. for 1 h. CombiFlash purification on a 12 g column eluted with a CH₂Cl₂ to 90:9:1 CH₂Cl₂:MeOH:2M NH₃ in MeOH gradient afforded the title compound as a tan solid. LC-MS m/z 846.4 (M+H)⁺, 1.71 min (ret time); mp 119-123° C.; analytical HPLC shows 97.2% purity, (ret time 11.996 min).

Example 86 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (0.706 g, 1.518 mmol), lithium hydroxide (0.0545 g, 2.28 mmol), THF (60 mL), and water (20 mL) were combined and stirred under argon at room temperature overnight. Solvents were pumped off, and the residue taken up in water (20 mL). The mixture was acidified with HCl until it appeared that no more precipitate was forming. The pH at this time was about 6. The solid was filtered off and washed 2× with a small volume of water. The material was dried under vacuum for two days until weight loss was minimal to give the title compound as a white solid. LC-MS m/z 451.8 (M+H)⁺, 1.48 min (ret time); mp 207-209° C.; analytical HPLC shows 100% purity, (ret time 10.177 min).

Example 87 3-Bromo-4-methylbenzamide

3-Bromo-4-methylbenzoic acid (5 g, 23.25 mmol) was suspended in CH₂Cl₂ (100 mL) and stirred under argon at room temperature. Oxalyl chloride (5.9 g, 46.5 mmol) was added followed by DMF (20 μL). Gas evolution began, and the mixture was stirred for 2 days during which time complete solution occurred. The solvents were pumped off and toluene was added and stripped off to remove excess oxalyl chloride. The residue was taken up in EtOAc and added to concentrated ammonium hydroxide (20 mL). This was stirred for thirty min. The phases were separated and the organic washed 1× with brine dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was crystallized from EtOAc/hexane and dried under vacuum to afford the title compound as a white crystalline solid. LC-MS m/z 213.8 (M+H)⁺, 1.41 min (ret time); mp 178-179° C.

Example 88 [(3-Bromo-4-methylphenyl)methyl]amine

To 3-bromo-4-methylbenzamide (2.14 g, 10 mmol) in THF (10 mL) was added borane dimethyl sulfide complex (2 mL, 20 mmol) at 0° C. The mixture was then heated to 50° C. for 16 h. Additional borane dimethyl sulfide complex (1 mL, 10 mmol) was added and heating continued at 60° C. for an additional 5 days. The reaction was cooled to room temperature and ethanol was cautiously added. When bubbling ceased, 1N HCl was added until pH was ˜2. The mixture was stirred at 50° C. for 4 h. The mixture was partitioned between EtOAc and water. The aqueous was washed 3× with EtOAc. The aqueous was then adjusted to pH 10 with 2N NaOH and extracted 3× with EtOAc. The combined organics phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound. LC-MS m/z 199.8 (M+H)⁺, 1.01 min (ret time).

Example 89 N-[(3-Bromo-4-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.451 g, 1 mmol), [(3-bromo-4-methylphenyl)methyl]amine (0.2 g, 1 mmol, HBTU (0.379 g, 1 mmol), and TEA (0.278 mL, 2 mmol) were combined in CH₂Cl₂ (25 mL). The mixture was stirred overnight at room temperature. Aqueous workup gave the crude product which was CombiFlashed on a 40 g column eluted with 0-10% MeOH/CH₂Cl₂ to give the title compound as a white solid. LC-MS m/z 633.8 (M+H)⁺, 1.85 min (ret time); mp 175-176° C.; analytical HPLC shows 97.5% purity, (ret time 13.568 min).

Example 90 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-[(3-Bromo-4-methylphenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.352 g, 0.556 mmol), (3-formylphenyl)boronic acid (0.108 g, 0.722 mmol), potassium carbonate (0.23 g, 1.67 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.0324 g, 0.028 mmol) were combined in dioxane (3 mL) and water (1 mL). The mixture was microwaved at 150° C. for 30 min. The reaction mixture was diluted with EtOAc and washed 3× with H₂O. The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified using CombiFlash on a 40 g column eluted with 0-10% MeOH/CH₂Cl₂ to give the title compound as a white solid. LC-MS m/z 659.1 (M+H)⁺, 1.93 min (ret time); mp 114-117° C.; analytical HPLC shows 96.3% purity, (ret time 13.675 min).

Example 91 {[3-Bromo-4-(methyloxy)phenyl]methyl}amine

3-Bromo-4-(methyloxy)benzonitrile (2.12 g, 10 mmol), THF (30 mL), and 1.5 M borane in THF (30 mL, 45 mmols) were combined and stirred under argon at reflux. The additional 1.5 M borane in THF (30 mL, 45 mmols) and reflux continued. THF (30 mL), and 1.5 M borane in THF (30 mL, 45 mmols) was again added and the mixture refluxed for a total of ten days to drive the reaction to completion. The reaction was worked up by the cautious addition of ethanol followed by 1N HCl until the pH was 2. This mixture was then heated to 50° C. for 4 h. The solvents were pumped off, and the residue partitioned between EtOAc and water. The aqueous phase was washed 3× with EtOAc and adjusted to pH 10 by the addition of 2.5 N NaOH. The aqueous phase was extracted 3× with EtOAc. The combined organic phases were dried over anhydrous Na₂SO₄, filtered and concentrated to give the title compound. LC-MS m/z 198.7 (M−NH₂), 0.99 min (ret time).

Example 92 N-{[3-Bromo-4-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.564 g, 1.25 mmol), {[3-bromo-4-(methyloxy)phenyl]methyl}amine (0.27 g, 1.25 mmol, HBTU (0.474 g, 1.25 mmol), and TEA (0.348 mL, 2.5 mmol) were combined in CH₂Cl₂ (30 mL). The mixture was stirred overnight at room temperature. The solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried and concentrated to give the crude product which was purified using CombiFlash on a 40 g column eluted with 0-10% MeOH/CH₂Cl₂ to afford the title compound as a white solid. LC-MS m/z 649.5 (M+H)⁺, 1.76 min (ret time); mp 130-132° C.; analytical HPLC shows 100% purity, (ret time 12.776 min).

Example 93 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[3-Bromo-4-(methyloxy)phenyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.64 g, 0.99 mmol), (3-formylphenyl)boronic acid (0.193 g, 1.29 mmol), potassium carbonate (0.41 g, 2.97 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.058 g, 0.05 mmol) were combined in dioxane (6 mL) and water (2 mL). The mixture was microwaved at 150° C. for 30 min. The reaction mixture was diluted with EtOAc and washed 3× with H₂O. The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified using CombiFlash on a 40 g column eluted with 0-10% MeOH/CH₂Cl₂ to give the title compound as a white solid. LC-MS m/z 675.1 (M+H)⁺, 1.85 min (ret time); mp 118-121° C.; analytical HPLC shows 100% purity, (ret time 13.204 min).

Example 94 3-Bromo-4-chlorobenzamide

3-Bromo-4-chlorobenzoic acid (2.35 g, 10 mmol) was suspended in CH₂Cl₂ (50 mL) and stirred under argon at room temperature. Oxalyl chloride (2.53 g, 20 mmol) was added followed by DMF (10 μL). Gas evolution began, and the mixture was stirred until gas evolution ceased. The solvents were pumped off and toluene was added and stripped off to remove excess oxalyl chloride. The residue was taken up in EtOAc and added to concentrated ammonium hydroxide (10 mL). This was stirred for thirty min. The phases were separated and the organic washed 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was crystallized from EtOAc/hexane to give the title compound as a white crystalline solid. LC-MS m/z 233.7 (M+H)⁺, 1.54 min (ret time); mp 146-147° C.; analytical HPLC shows 100% purity, (ret time 11.835 min).

Example 95 [(3-Bromo-4-chlorophenyl)methyl]amine

To 3-bromo-4-chlorobenzamide (1.6 g, 6.8 mmol) in THF (10 mL) was added borane dimethyl sulfide complex (1.36 mL, 13.6 mmol) at room temperature. The mixture was then heated to 60° C. for 8 days. The solvent was pumped off and the reaction cautiously quenched with ethanol. When bubbling ceased, 1N HCl was added until pH was ˜2. The mixture was stirred at 50° C. for 4 h. The mixture was partitioned between EtOAc and water. The aqueous phase was washed 3× with EtoAC. The aqueous was then adjusted to pH 10 with 2N NaOH and extracted 3× with EtOAc. The combined organics phases were washed with brine (1×) and dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a clear oil. LC-MS m/z 219.6 (M+H)⁺, 1.41 min (ret time).

Example 96 N-[(3-Bromo-4-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.564 g, 1.25 mmol), [(3-bromo-4-chlorophenyl)methyl]amine (0.275 g, 1.25 mmol, HBTU (0.474 g, 1.25 mmol), and TEA (0.348 mL, 2.5 mmol) were combined in CH₂Cl₂ (30 mL). The mixture was stirred overnight at room temperature. The reaction mixture was concentrated to dryness. Residue taken up in EtOAc, washed with 1 N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. It was purified further using CombiFlash on a 40 g column eluted with 0-10% MeOH/CH₂Cl₂ to give the title compound as a white solid. LC-MS m/z 652.9 (M+H)⁺, 1.91 min (ret time); mp 115-117° C.; analytical HPLC shows 98.4% purity, (ret time 13.689 min).

Example 97 N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(3-Bromo-4-chlorophenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.065 g, 0.1 mmol), (3-formylphenyl)boronic acid (0.0135 g, 0.09 mmol), potassium carbonate (0.041 g, 0.3 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.0058 g, 0.005 mmol) were combined in dioxane (1.5 mL) and water (0.5 mL). The mixture was microwaved at 100° C. for 15 min. Some starting material remained, so the mixture was microwaved again at 100° C. for 15 min. Again some starting material remained. Additional (3-formylphenyl)boronic acid (0.003 g, 0.02 mmol) was added, and the resulting mixture was microwaved again at 100° C. for 15 min. The reaction mixture was diluted with EtOAc and washed with H₂O and with brine. The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by CombiFlash on a 12 g column eluted with 0-10% MeOH/CH₂Cl₂ to give the title compound as a white solid. LC-MS m/z 679.0 (M+H)⁺, 1.84 min (ret time); mp 131-134° C.; analytical HPLC shows 97.2% purity, (ret time 13.823 min).

Example 98 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoate

3-Methyl-5-[(methyloxy)carbonyl]benzoic acid (2.91 g, 15.00 mmol) was partially dissolved in dichloromethane (DCM) (500 mL) with stirring under argon at room temperature. HBTU (5.69 g, 15.00 mmol) was added followed by 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine hydrochloride (5.10 g, 15 mmol), and then TEA (4.18 mL, 30.0 mmol). The mixture was stirred under argon overnight. The solvent was concentrated and the residue partitioned between EtOAc (300 mL) and water (100 mL). The organic phase was washed with water (3×100 mL), brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. This was purified by CombiFlash on a 120 g silica column eluted with 35-100% EtOAc in hexane. The purified product was taken up in 300 mL of EtOAc, and washed with 50 mL of 1N NaOH, and then with water (3×100 mL), and 50 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the title compound as a white solid. LC-MS m/z 480.0 (M+H)⁺, 0.81 min (ret time); mp 89-93° C.; analytical HPLC shows 95.4% purity, (ret time 12.251 min).

Example 99 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic Acid

Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoate (3.49 g, 7.28 mmol) was dissolved in tetrahydrofuran (THF) (90 ml) and water (30.0 ml) was added. Lithium hydroxide (0.458 g, 10.92 mmol) was added and the mixture stirred under argon at room temperature overnight. The THF was rotavaped off, and the aqueous residue was adjusted to ˜pH of 6 with 1N HCL. A white solid formed and was filtered and washed 2× with water (10 mL). This solid was dried at 50° C. in a vacuum oven for 6 h to give the title compound as a white solid. LC-MS m/z 466.0 (M+H)⁺, 0.73 min (ret time); mp 236-239° C.; analytical HPLC shows 97.3% purity, (ret time 10.841 min).

Example 100 N-{[3-Bromo-4-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic acid (0.931 g, 2 mmol) was partially dissolved in the dichloromethane (DCM) (50 mL), and then HBTU (0.758 g, 2.000 mmol) was added followed sequentially by {[3-bromo-4-(methyloxy)phenyl]methyl}amine (0.432 g, 2.000 mmol) and TEA (0.558 mL, 4.00 mmol). The mixture was allowed to stir under argon overnight at room temperature. The solvent was removed on a rotavap to give the crude product. The residue was taken up in EtOAc (100 mL) and washed 3× with water (50 mL), 1× with brine, dried over anhydrous Na₂SO₄ filtered and the solvent concentrated. The residue was taken up in methylene chloride and absorbed on Isolute® Sorbent and purified by CombiFlash on an 80 g silica column eluted with 0-10% DCM/MeOH to give the title compound as a white solid. LC-MS m/z 663.0 (M+H)⁺, 0.91 min (ret time); mp 123-126° C.; analytical HPLC shows 97.6% purity, (ret time 13.411 min).

Example 101 3-Bromo-5-chlorobenzamide

3-Bromo-5-chlorobenzoic acid (2.35 g, 10 mmol) was suspended in CH₂Cl₂ (50 mL) and stirred under argon at room temperature. Oxalyl chloride (2.53 g, 20 mmol) was added followed by DMF (10 μL), and the mixture stirred overnight. The solvents were pumped off. The residue was taken up in EtOAc and added to concentrated ammonium hydroxide (10 mL). This was stirred for thirty min. The phases were separated and the organic layer was washed 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was crystallized from EtOAc/hexane to give the title compound as a white crystalline solid. LC-MS m/z 233.7 (M+H)⁺, 1.57 min (ret time); analytical HPLC shows 96.5% purity, (ret time 12.131 min).

Example 102 [(3-Bromo-5-chlorophenyl)methyl]amine

To 3-bromo-5-chlorobenzamide (1.6 g, 6.8 mmol) in THF (10 mL) was added borane dimethyl sulfide complex (1.36 mL, 13.6 mmol) at room temperature. The mixture was then heated to 60° C. for 7 days. The solvent was pumped off and the reaction cautiously quenched with ethanol. When bubbling ceased, 1N HCl was added until pH was ˜2. The mixture was stirred at 50° C. for 4 h. The mixture was partitioned between EtOAc and water. The aqueous was washed 3× with ethyl acetate. The aqueous was then adjusted to pH 10 with 2N NaOH and extracted 3× with EtOAc. The combined organics phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a clear oil. LC-MS m/z 219.7 (M+H)⁺, 1.42 min (ret time).

Example 103 N-[(3-Bromo-5-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (677 mg, 1.5 mmol) was suspended in dichloromethane (DCM) (50 ml), and HBTU (569 mg, 1.5 mmol) was added, followed sequentially by 3-bromo-5-chlorobenzylamine (331 mg, 1.5 mmol), and TEA (0.418 mL, 3.0 mmol). The reaction mixture was stirred overnight under argon at room temperature to give a clear-colorless solution. The solvent was removed on a rotovap and the residue was partitioned between ethyl acetate (100 mL) and 1N NaOH (20 mL). The phases were separated and the organic phase was washed 2× with 1N NaOH (20 mL), and then 1× with brine (20 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered and concentrated to give the crude product. The crude product was taken up in dichloromethane and absorbed on Isolute® Sorbent and purified by CombiFlash on a 40 g silica column eluted with 0-10% DCM/MeOH to give the title compound as white solid. LC-MS m/z 652.9 (M+H)⁺, 2.08 min (ret time); mp 109-112° C.; analytical HPLC shows 97.6% purity, (ret time 13.921 min).

Example 104 N-[(5-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(3-Bromo-5-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (300 mg, 0.459 mmol), (3-formylphenyl)boronic acid (68.8 mg, 0.459 mmol), potassium carbonate (190 mg, 1.376 mmol), and tetrakis(triphenylphosphine)palladium(0) (26.5 mg, 0.023 mmol) were placed in a 2-5 mL Biotage microwave vial along with a stir bar. Then 1,4-dioxane (3 mL) and water (1 mL) were added. The vial was capped and microwaved for 15 min at normal power in the SmithCreator microwave at 100° C. The reaction mixture was partitioned between ethyl acetate (50 mL) and water (10 mL). The organic phase was washed 3× with water (10 mL), 1× with brine (10 mL), and dried over anhydrous Na₂SO₄. The crude product was taken up in dichloromethane and absorbed on Isolute® Sorbent and purified using CombiFlash on a 40 g silica column eluted with 0-10% DCM/MeOH to afford the title compound as a pale yellow solid. LC-MS m/z 679.0 (M+H)⁺, 2.21 min (ret time); mp 123-127° C.; analytical HPLC shows 96.2% purity, (ret time 14.171 min).

Example 105 1,6-Diethyl-5-[(methylamino)methyl]-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine

To the solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.303 g, 1.0 mmol) in THF (1 mL) was added Boc₂O (0.229 g, 1.05 mmol) with stirring at room temperature for 30 min whereupon LAH (5.0 mL, 1.0 M in THF) was added and the mixture heated with a microwave at 100° C. for 30 min. The reaction was then quenched with Na₂SO₄ (sat. aq.), filtered, dried over Na₂SO₄, filtered, concentrated to afford the title compound 0.252 g (79%). LC-MS m/z 318 (M+H)⁺.

Example 106 {3-[(4-{[(1,1-Dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]phenyl}boronic Acid

To a solution of (3-formylphenyl)boronic acid (3.0 g, 20.0 mmol) in DCM (100 mL) was added 1,1-dimethylethyl 1-piperazinecarboxylate (3.91 g, 21.0 mmol), NaBH(OAc)₃ (6.36 g, 30.0 mmol) and this mixture was stirred at room temperature for 17 h. The organic layer was diluted with EtOAc (100 mL), which was washed with H₂O (30 mL), then dried over Na₂SO₄, filtered and concentrated to afford the title compound 7.72 g (quantitative). LC-MS m/z 321 (M+H)⁺, 1.49 min (ret time).

Example 107 1,1-Dimethylethyl 4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate

To two vials which each contained a solution of [(3-bromo-4-fluorophenyl)methyl]amine hydrochloric salt (0.601 g, 2.5 mmol) in p-dioxane/H₂O (15/5 mL) was added {3-[(4-{[(1,1-dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]phenyl}boronic acid (1.2 g, 3.75 mmol), Pd(PPh₃)₄ (145 mg, 0.125 mmol), and K₂CO₃ (1.38 g, 10 mmol). The mixture was heated with a microwave at 150° C. for 15 min. The organic layer of both vials was separated, combined, concentrated and purified by CombiFlash chromatograph to afford the title compound 1.98 g (99%). LC-MS m/z 400 (M+H)⁺.

Example 108 1,1-Dimethylethyl 4-({5′-[({[3-(chloromethyl)phenyl]carbonyl}amino) methyl]-2′-fluoro-3-biphenylyl}methyl)-1-piperazinecarboxylate

To a solution of 1,1-dimethylethyl 4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.32 g, 0.80 mmol) in p-dioxane/DCM (4/4 mL) was added 3-(chloromethyl)benzoic acid (0.12 g, 0.88 mmol), EDC (0.23 g, 1.2 mmol), HOBt (0.011 g, 0.08 mmol), and Et₃N (0.112 mL, 0.8 mmol). The mixture was stirred at room temperature for 50 min before was the reaction was quenched with H₂O (3 mL). The organic layer was separated, dried over Na₂SO₄, filtered, concentrated and purified by CombiFlash chromatograph to afford the title compound 0.216 g (49%). LC-MS m/z 552 (M+H)⁺.

Example 109 1,1-Dimethylethyl 4-{[5′-({[(3-{[{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}(methyl)amino]methyl}phenyl)carbonyl]amino}methyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate

To a solution of 1,6-diethyl-5-[(methylamino)methyl]-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (0.0432 g, 0.136 mmol) in DMF (1.0 mL) was added 1,1-dimethylethyl 4-({5′-[({[3-(chloromethyl)phenyl]carbonyl}amino)methyl]-2′-fluoro-3-biphenylyl}methyl)-1-piperazinecarboxylate (0.075 g, 0.136 mmol), and K₂CO₃ (0.0188 g, 0.136 mmol). This mixture was stirred at room temperature for 18 h after which was added NaI (0.0204 g, 0.136 mmol) and stirring continued for another 25 h. Then the mixture was diluted with EtOAc (5 mL), washed with H₂O (2×3 mL), brine (3 mL), concentrated, and purified by CombiFlash chromatograph to afford the title compound 0.055 g (49%). LC-MS m/z 833 (M+H)⁺.

Example 110 1,1-Dimethylethyl 4-[(2′-fluoro-5′-{[({3-[(methyloxy)carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-1-piperazinecarboxylate

To a solution of 1,1-dimethylethyl 4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.200 g, 0.5 mmol) in CH₂Cl₂ (5 mL) was added 3-[(methyloxy)carbonyl]benzoic acid (0.0901 g, 0.5 mmol), HBTU (0.19 g, 0.5 mmol), and Et₃N (0.0703 mL, 0.5 mmol). The mixture was stirred at room temperature for 18 h and then quenched with H₂O. The organic layer was separated, concentrated and purified by CombiFlash chromatograph to afford the title compound 0.26 g (93%). LC-MS m/z 562 (M+H)⁺.

Example 111 3-{[({3′-[(4-{[(1,1-Dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)amino]carbonyl}benzoic Acid

To a solution of 1,1-dimethylethyl 4-[(2′-fluoro-5′-{[({3-[(methyloxy)carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-1-piperazinecarboxylate (0.26 g, 0.463 mmol) in MeOH/H₂O (2/1 mL) was added LiOH.H₂O (0.0971 g, 2.32 mmol). The mixture was heated with a microwave at 80° C. for 30 min. Then HCl was added to adjust the pH to ˜5. Then solvents were removed. The residue was extracted with DCM to afford the title compound 0.256 g (100%). LC-MS m/z 548 (M+H)⁺.

Example 112 1,1-Dimethylethyl 4-{[5′-({[(3-{[{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}(methyl)amino]carbonyl}phenyl)carbonyl]amino}methyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate

To a solution of 1,6-diethyl-5-[(methylamino)methyl]-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (0.074 g, 0.233 mmol) in DMF (2.3 mL) was added 3-{[({3′-[(4-{[(1,1-dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)amino]carbonyl}benzoic acid (0.127 g, 0.233 mmol), HBTU (0.133 g, 0.35 mmol), and Et₃N (0.0655 mL, 0.466 mmol). The mixture was stirred at room temperature for 19 h when it was quenched with H₂O. The organic layer was separated, concentrated and purified by CombiFlash chromatograph to afford the title compound 0.0565 g (29%). LC-MS m/z 847 (M+H)⁺.

Example 113 1,1-Dimethylethyl [(3-bromo-4-fluorophenyl)methyl]carbamate

To a solution of [(3-bromo-4-fluorophenyl)methyl]amine hydrochloric salt (0.64 g, 2.0 mmol) in THF (10 mL) was added NaOH (2 mL, 1.0 M, 2.0 mmol). This mixture was stirred for 10 min and then Boc₂O (0.523 g, 2.4 mmol) was added, and the mixture was stirred for another 2 h. The organic layer was then separated, dried, filtered, concentrated and purified by CombiFlash chromatograph to afford the title compound 0.66 g (quantitative). LC-MS m/z 609 (2M+H)⁺.

Example 114 [(3-Bromo-4-fluorophenyl)methyl]methylamine

To a solution of 1,1-dimethylethyl [(3-bromo-4-fluorophenyl)methyl]carbamate (0.755 g, 2.48 mmol) in THF (1 mL) was added BH₃.THF (12.5 mL, 1.0 M in THF). The mixture was heated with a microwave at 80° C. for 30 min twice. The reaction was then quenched with HCl (10 mL, 1 N) slowly, after which the mixture was stirred for 2 h at room temperature, basified with NaHCO₃ to pH ˜9, and extracted with EtOAc (50+20 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, concentrated to afford the title compound 0.52 g (96%). LC-MS m/z 218 (M+H)⁺.

Example 115 1,1-Dimethylethyl 4-({2′-fluoro-5′-[(methylamino)methyl]-3-biphenylyl}methyl)-1-piperazine carboxylate

To a solution of [(3-bromo-4-fluorophenyl)methyl]methylamine (0.52 g, 2.39 mmol) in p-dioxane/H₂O (15/5 mL) was added {3-[(4-{[(1,1-dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]phenyl}boronic acid (1.15 g, 3.60 mmol), Pd(PPh₃)₄ (139 mg, 0.12 mmol), and K₂CO₃ (1.33 g, 9.6 mmol). The mixture was heated with a microwave at 150° C. for 15 min. The organic layer was separated, concentrated. The residue was redissolved in EtOAc (˜70 mL), which was washed with H₂O (20 mL), brine (20 mL), dried over Na₂SO₄, filtered, concentrated and purified by CombiFlash chromatograph to afford the title compound 0.65 g (66%). LC-MS m/z 414 (M+H)⁺.

Example 116 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

To a solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.303 g, 1.0 mmol) in CH₂Cl₂ (10 mL) was added 3-[(methyloxy)carbonyl]benzoic acid (0.198 g, 1.1 mmol), HBTU (0.569 g, 1.5 mmol), and Et₃N (0.281 mL, 2.0 mmol). The mixture was stirred at room temperature for 22 h. The reaction was quenched with H₂O (1 mL). The organic layer was separated, directly load onto silica gel cartridge and purified by CombiFlash chromatograph to afford the title compound 0.29 g (55%). LC-MS m/z 466 (M+H)⁺.

Example 117 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino) carbonyl]benzoic Acid

To the solution of methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (0.29 g, 0.623 mmol) in MeOH/H₂O (2/1 mL) was added LiOH.H₂O (0.131 g, 3.12 mmol). The mixture was heated with a microwave at 80° C. for 30 min. Then HCl (1 N) was added to adjust the about pH 4˜5. The solvent was removed. The residue was washed with H₂O and a little EtOAc and dried under vacuum to afford the title compound 0.20 g (70%). LC-MS m/z 452 (M+H)⁺.

Example 118 1,1-Dimethylethyl 4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)(methyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-1-piperidinecarboxylate

To a solution of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridine-5-yl]methyl}amino) carbonyl]benzoic acid (0.197 g, 0.436 mmol) in DMF (4.5 mL) was added 1,1-dimethylethyl-4-({2′-fluoro-5′-[(methylamino)methyl]-3-biphenylyl}methyl)-1-piperazine carboxylate (0.18 g, 0.436 mmol), HBTU (0.248 g, 0.654 mmol), and Et₃N (0.122 mL, 0.872 mmol). The resulting mixture was stirred at room temperature for 66 h. The reaction was diluted with EtOAc (50 mL), washed with H₂O (20+2×10 mL), brine (20 mL), dried over Na₂SO₄. The dried solution was filtered, concentrated, and purified by CombiFlash chromatograph to afford 0.089 g (24%) of the title compound. LC-MS m/z 847 (M+H)⁺.

Example 119 3-(Chloromethyl)-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}benzamide

To a solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.303 g, 1.0 mmol) in CH₂Cl₂ (10 mL) was added 3-(chloromethyl)benzoic acid (0.188 g, 1.1 mmol), EDC (0.288 g, 1.5 mmol), HOBt (0.0135 g, 0.1 mmol), and Et₃N (0.141 mL, 1.0 mmol). This mixture was stirred at room temperature for 1 h, then washed with H₂O (1 mL) and purified by CombiFlash chromatograph to afford the title compound 0.285 g (63%). LC-MS m/z 456 (M+H)⁺.

Example 120 3-{[[(3-Bromo-4-fluorophenyl)methyl](methyl)amino]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}benzamide

To a solution of 3-(chloromethyl)-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}benzamide (0.284 g, 0.623 mmol) in DMF (2.5 mL) was added [(3-bromo-4-fluorophenyl)methyl]methylamine (0.436 g, 2.0 mmol). This mixture was stirred at room temperature for 66 h. Then EtOAc (50 mL) was added and that solution washed with H₂O (7.5+2×5 mL), brine (5 mL), dried over Na₂SO₄. The dried organic solution was filtered, concentrated and purified by CombiFlash chromatograph to afford the title compound 0.371 g (93%). LC-MS m/z 637 (M+H)⁺.

Example 121 1,1-Dimethylethyl 4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino) carbonyl]phenyl}methyl)(methyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-1-piperazinecarboxylate

To a solution of 3-{[[(3-bromo-4-fluorophenyl)methyl](methyl)amino]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}benzamide (0.37 g, 0.58 mmol) in p-dioxane/H₂O (4.5/1.5 mL) was added {3-[(4-{[(1,1-dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]phenyl}boronic acid (0.279 g, 0.87 mmol), Pd(PPh₃)₄ (33.6 mg, 0.029 mmol), and K₂CO₃ (0.32 g, 2.32 mmol). The mixture was heated with a microwave at 150° C. for 15 min. The organic layer was separated and concentrated. The residue was redissolved in EtOAc, washed with H₂O, brine, dried over Na₂SO₄, filtered, concentrated and the concentrate purified by CombiFlash chromatograph to afford the title compound 0.396 g (82%). LC-MS m/z 833 (M+H)⁺.

Example 122 5-(Chloromethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine

To a solution of thionyl chloride (1.46 mL, 20.0 mmol) was added [1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methanol (0.609 g, 2.0 mmol) slowly. The mixture was stirred at room temperature for 30 min and then was concentrated on rotavap under vacuum. The residue was twice taken up in DCM (5 mL) and concentrated on rotavap to afford the title compound 0.386 g (60%). LC-MS m/z 319 (M+H)⁺ [molecular ion of the compound with methoxy from LC-MS solvent methanol where OMe substituted chloro group of the original product].

Example 123 3-(Hydroxymethyl)benzoic Acid

To 3-[(methyloxy)carbonyl]benzoic acid (0.18 g, 1.0 mmol) was added lithium triethyl borohydride (3.0 mL, 1.0 M in THF, 3.0 mmol) while stirring. The mixture was stirred for 3 h and then quenched with NH₄Cl (sat.) and acidified with HCl (aq.). This solution was extracted with EtOAc, and the extract was dried over Na₂SO₄, filtered and concentrated to afford the title compound 0.136 g (89%). LC-MS m/z 153 (M+H)⁺.

Example 124 1,1-Dimethylethyl 4-({2′-fluoro-5′-[({[3-(hydroxymethyl)phenyl]carbonyl}amino)methyl]-3-biphenylyl}methyl)-1-piperazinecarboxylate

To a solution of 1,1-dimethylethyl 4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate (100 mg, 0.25 mmol) in DCM (2.5 mL) was added 3-(hydroxymethyl)benzoic acid (38 mg, 0.25 mmol), HBTU (0.104 g, 0.275 mmol), and Et₃N (0.0351 mL, 0.25 mmol). The mixture was stirred at room temperature for 21 h. The reaction was quenched with H₂O. The organic layer was separated, directly load onto silica gel cartridge and purified by CombiFlash chromatograph to afford the title compound 0.122 g (91%). LC-MS m/z 534 (M+H)⁺.

Example 125 Methyl 3-{[[(3-bromo-4-fluorophenyl)methyl](methyl)amino]carbonyl}benzoate

To a solution of [(3-bromo-4-fluorophenyl)methyl]methylamine (0.355 g, 1.63 mmol) in THF (16 mL) was added 3-[(methyloxy)carbonyl]benzoic acid (0.294 g, 1.63 mmol), HBTU (0.929 g, 2.45 mmol), Et₃N (0.458 mL, 3.26 mmol) and stirred at room temperature for 65 h, and then quenched with H₂O. The organic layer was loaded onto silica gel cartridge and purified by CombiFlash chromatograph to afford the title compound 0.419 g (68%). LC-MS m/z 380 (M+H)⁺.

Example 126 1,1-Dimethylethyl (2S)-4-[(2′-fluoro-5′-{[methyl({3-[(methyloxy) carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate

To a solution of methyl 3-{[[(3-bromo-4-fluorophenyl)methyl](methyl)amino]carbonyl}benzoate (0.20 g, 0.526 mmol) in p-dioxane/H₂O (4.5/1.5 mL) was added {3-[((3S)-4-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]-phenyl}boronic acid (0.264 g, 0.789 mmol), Pd(PPh₃)₄ (0.030 g, 0.026 mmol), and K₂CO₃ (0.29 mL, 2.1 mmol). The mixture was heated with z microwave at 130° C. for 15 min. The mixture was then filtered, and the filtrate concentrated and purified by CombiFlash chromatograph to afford the title compound 0.215 g (69%). LC-MS m/z 590 (M+H)⁺.

Example 127 3-{[({3′-[((3S)-4-{[(1,1-Dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)(methyl)amino]carbonyl}benzoic Acid

To a solution of 1,1-dimethylethyl (2S)-4-[(2′-fluoro-5′-{[methyl({3-[(methyloxy)carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate (0.215 g, 0.365 mmol) in MeOH/H₂O (1.0/0.5 mL) was added LiOH.H₂O (0.077 g, 1.83 mmol). The mixture was heated with a microwave at 80° C. for 30 min. Then it was acidified with HOAc. This mixture was extracted with DCM which was dried over Na₂SO₄, filtered, concentrated and purified using a Gilson HPLC to afford the title compound 0.0639 g (30%). LC-MS m/z 576 (M+H)⁺.

Example 128 Methyl 3-{[[(6-fluoro-3′-formyl-3-biphenylyl)methyl](methyl)amino]carbonyl}benzoate

To a solution of methyl 3-{[[(3-bromo-4-fluorophenyl)methyl]-(methyl)amino]carbonyl}benzoate (0.20 g, 0.526 mmol) in p-dioxane/H₂O (4.5/1.5 mL) was added (4-formylphenyl)boronic acid (0.118 g, 0.789 mmol), Pd(PPh₃)₄ (0.030 g, 0.026 mmol), and K₂CO₃ (0.29 mL, 2.1 mmol). The mixture was heated in a microwave at 130° C. for 15 min. It was then filtered, concentrated and the concentrate purified by CombiFlash chromatograph to afford the title compound 0.184 g (86%). LC-MS m/z 406 (M+H)⁺.

Example 129 Methyl 3-{[({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)(methyl)amino]carbonyl}benzoate

To a solution of methyl 3-{[[(6-fluoro-3′-formyl-3-biphenylyl)methyl](methyl)amino]carbonyl}benzoate (0.18 g, 0.444 mmol) in DCM (4.4 mL) was added 1-methylpiperazine (0.0542 mL, 0.488 mmol), and NaBH(OAc)₃ (0.141 g, 0.666 mmol). The mixture was stirred at room temperature for 17 h and then the reaction was quenched with H₂O (˜1 mL). The organic layer was separated, dried over Na₂SO₄ and purified by CombiFlash chromatograph to afford the title compound 0.202 g (93%). LC-MS m/z 490 (M+H)⁺.

Example 130 3-{[({6-Fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)(methyl)amino]carbonyl}benzoic Acid

To a solution of methyl 3-{[({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)(methyl)amino]carbonyl}benzoate (0.20 g, 0.409 mmol) in MeOH/H₂O (1.2/0.6 mL) was added LiOH.H₂O (0.086 g, 2.05 mmol). The mixture was heated in a microwave at 80° C. for 30 min following which it was acidified with HCl, concentrated and the concentrate purified using a Gilson HPLC to afford the title compound 0.221 g (quantitative). LC-MS m/z 476 (M+H)⁺.

Example 131 Methyl 4-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

To a solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (1.02 g, 3.0 mmol) in DCM (30 mL) was added 4-[(methyloxy)carbonyl]benzoic acid (0.541 g, 3.0 mmol), HBTU (1.37 g, 3.6 mmol), and Et₃N (0.843 mL, 6.0 mmol) This mixture was stirred at room temperature for 23 h. It was diluted with DCM (30 mL), which was washed with H₂O (30 mL) and dried over Na₂SO₄, filtered, concentrated and the concentrate purified by CombiFlash chromatograph to afford the title compound 1.40 g (quantitative). LC-MS m/z 466 (M+H)⁺.

Example 132 4-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino) carbonyl]benzoic Acid

To a solution of 3-{[({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl) (methyl)amino]carbonyl}benzoic acid (1.40 g, 3.0 mmol) in MeOH/H₂O (9.0/4.5 mL) was added LiOH.H₂O (0.634 g, 15.0 mmol). The mixture was heated in a microwave at 80° C. for 30 min and then filtered, concentrated, acidified with HCl. The acidic solution was extracted with EtOAc which was dried over Na₂SO₄, filtered, concentrated to afford the title compound 0.984 g (73%). LC-MS m/z 452 (M+H)⁺.

Example 133 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

A mixture of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (1.0 g, 2.94 mmol), 3-[(methyloxy)-carbonyl]benzoic acid (0.58 g, 3.24 mmol), HBTU (1.34 g, 3.53 mmol) and Et₃N (0.82 mL, 5.89 mmol) in DCM (30 mL) was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO₃ and the solution extracted with DCM twice. The combined organic layers were washed with water followed by a brine wash. The organic layer was dried over sodium sulfate, filtered, concentrated and purified using CombiFlash chromatography, eluting with 80% ethyl acetate in hexane. The product fractions were combined and concentrated to give methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)-carbonyl]benzoate as an oil (1.40 g, 99%). LC-MS m/z 467 (M+H)⁺.

Example 134 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

A mixture of methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (8.52 g, 18.3 mmol), LiOH (3.84 g, 91.5 mmol), water (30 mL) and methanol (60 mL) was stirred at room temperature overnight. The aqueous layer was acidified with 2 N HCl to pH around 1. Brine (30 mL) was added to the solution. White solid was precipitated out from the solution. The solid was filtered off and washed with water. The aqueous layer was extracted with ethyl acetate twice. The combined organic layers were concentrated under vacuum to give a solid. All solid was dissolved in DCM and combined into one batch. Solvent was concentrated to give crude residue. The crude residue was then triturated with ether to give 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid as a white solid (7.72 g, 86%). LC-MS m/z 452 (M+H)⁺.

Example 135 N-{[3-Bromo-5-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

A mixture of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (762 mg, 1.69 mmol), {[3-bromo-5-(methyloxy)phenyl]methyl}amine (529 mg, 1.86 mmol), HBTU (769 mg, 2.03 mmol) and Et₃N (0.47 mL, 3.38 mmol) in DCM was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were then washed with water followed by a brine wash. The organic layer was then concentrated under vacuum to give a crude residue. It was then purified using CombiFlash chromatography eluting with 0 to 10% methanol in DCM to obtain N-{[3-bromo-5-(methyloxy)phenyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a solid (1.05 g, 96%). LC-MS m/z 649 (M+H)⁺.

Example 136 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

A mixture of N-{[3-bromo-5-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (782 mg, 1.20 mmol), 3-formylphenyl boronic acid (234 mg, 1.57 mmol), Na₂CO₃ (383 mg, 3.61 mmol), Pd(PPh₃)₄ (69.6 mg, 0.06 mmol), and water (3 mL) in dioxane (9 mL) was degassed for 5 min. This mixture was then heated under microwave for 30 min at 150° C. It was quenched with water and then extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a 0.45 μM filter and then concentrated to give a crude residue. It was then purified with Combi Flash chromatography eluting with 0 to 10% MeOH in DCM. The product fractions were combined and concentrated under vacuum to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a solid (687 mg, 85%). LC-MS m/z 676 (M+H)⁺.

Example 137 N-[(3-Bromo-5-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

A mixture of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (1000 mg, 2.05 mmol), [(3-bromo-5-methylphenyl)methyl]amine (605 mg, 2.25 mmol), HBTU (932 mg, 2.45 mmol) and Et₃N (0.57 mL, 4.10 mmol) in DCM was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were then washed with water followed by a brine wash. The organic layer was then concentrated under vacuum to give a crude residue. It was then purified with CombiFlash chromatography, eluting with 0 to 10% methanol in DCM to obtain N-[(3-bromo-5-methylphenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as an oil (652 mg, 50%). LC-MS m/z 634 (M+H)⁺.

Example 138 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

A mixture of N-[(3-bromo-5-methylphenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (652 mg, 1.00 mmol), 3-formylphenyl Boronic acid (196 mg, 1.31 mmol), Na₂CO₃ (319 mg, 3.01 mmol), Pd(PPh₃)₄ (58 mg, 0.05 mmol), and water (2 mL) in dioxane (6 mL) was degassed for 5 min. and then heated in a microwave for 30 min at 150° C. The reaction was quenched with water and the mixture extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a pad of celite to get rid of Pd content and then concentrated to give a crude residue. It was then purified with Combi Flash chromatography eluting with 0 to 10% MeOH in DCM. The product fractions were combined and concentrated under vacuum to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a solid (654 mg, 71%). LC-MS m/z 660 (M+H)⁺.

Example 139 N-[(5-Bromo-2-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

To a clear solution of 1,1-dimethylethyl [(5-bromo-2-methylphenyl)methyl]carbamate (923 mg, 3.01 mmol) in DCM (10 mL) was added 4N HCl in dioxane (3 mL). Solid precipitated out from the solution after 30 min. The mixture was stirred at RT for another 1 h. It was then concentrated under vacuum to get rid of solvent. The resulting residue was re-dissolved in DCM. Et₃N (1.5 mL, 10.3 mmol) was added followed by 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)-carbonyl]benzoic acid (1000 mg, 2.05 mmol) and HBTU (933 mg, 2.46 mmol). The mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were then washed with water followed by the brine wash. The organic layer was then concentrated under vacuum to give a crude residue. It was then purified with CombiFlash chromatography, eluting with 0 to 10% methanol in DCM to obtain N-[(5-bromo-2-methylphenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a solid (1.4 g, 98%). LC-MS m/z 634 (M+H)⁺.

Example 140 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

A mixture of N-[(5-bromo-2-methylphenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (1000 mg, 1.58 mmol), 3-formylphenyl boronic acid (308 mg, 2.05 mmol), Na₂CO₃ (502 mg, 4.74 mmol), Pd(PPh₃)₄ (91 mg, 0.08 mmol), water (2 mL) in dioxane (6 mL) was degassed for 5 min. and then heated under microwave for 30 min at 150° C. The reaction was quenched with water and the mixture extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a pad of celite to get rid of Pd content and then concentrated to give a crude residue. It was then purified with Combi Flash chromatography, eluting with 0 to 10% MeOH in DCM. The product fractions were combined and concentrated under vacuum to give n-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a solid (678 mg, 65%). LC-MS m/z 660 (M+H)⁺.

Example 141 N-[(3-Bromo-5-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

To a clear solution of 1,1-dimethylethyl [(3-bromo-5-fluorophenyl)methyl]carbamate (280 mg, 1.11 mmol) 207310-29 in DCM (1 mL) was added 4N HCl in dioxane (1 mL). Solid precipitated out from the solution after 30 min. but the mixture was stirred at RT for another hour. It was then concentrated under vacuum to get rid of solvent. Then the residue was re-dissolved in DCM. Et₃N (0.5 mL, 3.07 mmol) was added followed by 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (300 mg, 0.62 mmol) and HBTU (311 mg, 3.07 mmol). The mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The organic layer was then washed with water followed by the brine wash. The organic layer was then concentrated under vacuum to give a crude residue. It was purified with CombiFlash chromatography, eluting with 0 to 10% methanol in DCM to obtain N-[(3-bromo-5-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a solid (307 mg, 78%). LC-MS m/z 637 (M+H)⁺.

Example 142 1,1-Dimethylethyl [(5-fluoro-3′-formyl-3-biphenylyl)methyl]carbamate

A mixture of 1,1-dimethylethyl [(3-bromo-5-fluorophenyl)methyl]carbamate (300 mg, 0.99 mmol), 3-formylphenyl boronic acid (194 mg, 1.30 mmol), Na₂CO₃ (316 mg, 2.98 mmol), Pd(PPh₃)₄ (58 mg, 0.05 mmol), and water (2 mL) in dioxane (6 mL) was degassed for 5 min. and then heated under microwave for 30 min at 150° C. The reaction was quenched with water and then extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a syringe filter to get rid of Pd content and then concentrated to give a crude residue. It was purified with Combi Flash chromatography, eluting with 40% ethyl acetate in hexane. The product fractions were combined and concentrated under vacuum to give 1,1-dimethylethyl [(5-fluoro-3′-formyl-3-biphenylyl)methyl]carbamate as an oil: LC-MS m/z 330 (M+H)⁺.

Example 143 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

To a clear solution of 1,1-dimethylethyl [(5-fluoro-3′-formyl-3-biphenylyl)methyl]carbamate (486 mg, 1.48 mmol) in DCM (2 mL) was added 4N HCl in dioxane (1 mL). Solid precipitated out from the solution after 30 min but stirring was continued at RT for another 1 h. The mixture was then concentrated under vacuum to get rid of solvent. The resulting residue was re-dissolved in DCM. Et₃N (1.14 mL, 8.20 mmol) was added followed by 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (400 mg, 0.82 mmol) and HBTU (830 mg, 8.20 mmol). The mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The organic layer was then washed with water followed by a brine wash. The organic layer was concentrated under vacuum to give a crude residue. It was purified with CombiFlash chromatography, eluting with 90% ethyl acetate in hexane to obtain N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (300 mg, 55%). LC-MS m/z 664 (M+H)⁺

Example 144 1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridine-5-carboxylic Acid

A mixture of ethyl 1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridine-5-carboxylate (2 g, 4.30 mmol), LiOH (901 mg, 21.48 mmol), water (4 mL) and methanol (8 mL) was heated at microwave for 20 min at 80° C. The reaction mixture was diluted with water and then washed with ethyl acetate to get rid of starting material. The aqueous layer was then acidified with 2N HCl, saturated with brine and then extracted with a mixture of DCM and IPA (3:1 ratio) twice. The combined organic layers were dried under vacuum to give a crude product. It was triturated with ether to give pure product 1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridine-5-carboxylic acid as a yellow solid (2.78 g, 95%). LC-MS m/z 319 (M+H)⁺.

Example 145 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

A mixture of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (2.0 g, 0.0059 mol) in DCM (60 mL) with 3-[(methyloxy)carbonyl]benzoic acid (1.16 g, 0.00648 mol) and HATU (2.68 g, 0.00706 mol) and TEA (1.19 g, 1.64 mL, 0.0118 mol) gave a yellow solution upon stirring overnight at RT. The solution was washed in turn with sat. aq. NaHCO₃, H₂O, and sat. aq. NaCl, then dried (Na₂SO₄), and concentrated to a viscous syrup/foam (3.68 g). This was purified on a ‘CombiFlash’ 40 g silica gel column (hexane-EtOAc) [20-100% EtOAc] to afford the title compound (3.02 g). LC-MS m/z 466.1 (M+H)⁺.

Example 146 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (3.02 g, 0.0064 mol) in MeOH (30 mL) was stirred with lithium hydroxide monohydrate (1.36 g, 0.032 mol) in H₂O (15 mL) overnight at room temperature. The solution was treated with 3N HCl to adjust the pH to about 2.0 and cooled in an ice bath. Solid was collected and dried (2.4 g). The filtrate was treated with sat. aq. NaCl and extracted with EtOAc (2×). The extract was dried (Na₂SO₄) and concentrated to a residue. It was triturated with ether to afford the title compound as a white solid (143 mg only slightly inferior to the main product by LC-MS). Total product 2.56 g (88.7%). LC-MS m/z 452.1 (M+H)⁺.

Example 147 N-[(3-Bromo-4-cyanophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (918 mg, 0.002 mol) and 4-(aminomethyl)-2-bromobenzonitrile HCl (545 mg, 0.0022 mol) in DCM (25 mL) was treated with HATU (912 mg, 0.0024 mol) followed by TEA (0.404 g, 0.56 mL, 0.004 mol). The reaction mixture was stirred overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃ and the layers separated. The aqueous phase was re-extracted with DCM (2×). The combined organic extracts were washed with H₂O, then sat. aq. NaCl, dried (Na₂SO₄), then the solvent was concentrated giving a residue (1.4 g). This was purified on the ‘CombiFlash’ using 40 g silica gel column [DCM-MeOH, 0-10%]. This afforded the title compound as a glassy, pale yellow foam (1.4 g). LC-MS m/z 645 (M+H)⁺.

Example 148 N-[(6-Cyano-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

A mixture of N-[(3-bromo-4-cyanophenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.775 g, 0.00120 mol) in dioxane (9 mL) and (3-formylphenyl)boronic acid (234.7 mg, 0.001565 mol) was treated with a solution of Na₂CO₃ (382.8 mg, 0.00361 mol) in H₂O (3 mL). The mixture was purged with N₂ for 10 min, then tetrakistriphenylphosphine palladium (0) (69 mg, 0.00060 mol) was added. The mixture was microwaved at 150° C. for 30 min. The reaction was quenched with H₂O and then extracted with EtOAc (2×). The combined organic extracts were washed with H₂O, then sat. aq. NaCl, dried (Na₂SO₄) and solvent concentrated to give a residue which was purified on a Chomatotrom™ Plate (2000μ) silica gel [DCM-MeOH, 0-6%] to afford the title compound [609 mg (76%)]. LC-MS m/z 671 (M+H)⁺.

Example 149 N-{[5-Bromo-2-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (918 mg, 0.002 mol) with 1-[5-bromo-2-(methyloxy)phenyl]methanamine, TFA salt (726 mg, 0.0022 mol) in DCM (25 mL) was treated with HATU (912 mg, 0.0024 mol) followed by TEA (0.404 g, 0.56 mL, 0.004 mol). One additional eq. of TEA was added. The reaction was stirred overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃. The layers were separated and the aqueous phase re-extracted with DCM (2×). Combined organic extracts were washed with H₂O, then sat. aq. NaCl, dried (Na₂SO₄), and then concentrated giving a residue (1.4 g). This was purified on the ‘CombiFlash’ using 40 g silica gel column [DCM-MeOH, 0-10%]. This afforded the title compound as a glassy, pale yellow foam [1.339 g (quant.)]. LC-MS m/z 651 (M+H)⁺.

Example 150 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

A mixture of N-[(3-bromo-4-cyanophenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.779 g, 0.00120 mol) in dioxane (9 mL) with (3-formylphenyl)boronic acid (234.7 mg, 0.001565 mol) was treated with a solution of Na₂CO₃ (382.8 mg, 0.00361 mol) in H₂O (3 mL). The mixture was purged with N₂ for 10 min, then Tetrakistriphenylphosphine palladium (0) (69 mg, 0.00060 mol) was added. The mixture was microwaved at 150° C. for 30 min. The reaction was quenched with H₂O, then the mixture was extracted with EtOAc (2×). The combined organic extracts were washed with H₂O, then sat. aq. NaCl, dried (Na₂SO₄) and concentrated giving a residue which was purified on a Chomatotrom™ Plate (2000μ) silica gel [DCM-MeOH, 0-6%] to afford the title compound (438 mg). However, there was loss of Br during the reaction and the des-bromo compound did not separate under the chromatographic conditions and the product 70% pure, with effectively 306 mg (38%) desired product obtained. LC-MS m/z 675.5 (M+H)⁺.

Example 151 N-[(3-Bromo-2-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (918 mg, 0.002 mol) and [(3-bromo-2-methylphenyl)methyl]amine hydrochloride (521 mg, 0.0022 mol) in DCM (25 mL) was treated with HATU (912 mg, 0.0024 mol) followed by TEA (0.404 g, 0.56 mL, 0.004 mol). The reaction was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction was quenched with sat. aq. NaHCO₃. The aqueous phase was re-extracted with DCM (2×). Combined organic extracts were washed with H₂O, then sat. aq. NaCl and dried (Na₂SO₄), then concentrated to give a residue (1.4 g). This was purified on the ‘CombiFlash’ using 40 g silica gel column [DCM-MeOH, 0-10%]. This afforded the title compound as a glassy, pale yellow foam [1.35 g (Quant.)]. LC-MS m/z 634.7 (M+H)⁺.

Example 152 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-2-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

A mixture of N-[(3-bromo-2-methylphenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.762 g, 0.00120 mol) in dioxane (9 mL) with (3-formylphenyl)boronic acid (234.7 mg, 0.001565 mol) was treated with a solution of Na₂CO₃ (382.8 mg, 0.00361 mol) in H₂O (3 mL). The mixture was purged with N₂ for 10 min, then tetrakistriphenylphosphine palladium (0)° (69 mg, 0.00060 mol) was added. The mixture was microwaved at 150° for 30 min. LC-MS showed that the reaction was complete. The reaction was quenched with H₂O, then extracted with EtOAc (2×). Combined organic extracts were washed with H₂O, then sat. aq. NaCl and dried (Na₂SO₄) and concentrated to give a residue which was purified on a Chomatotrom™ Plate (2000μ) silica gel [DCM-MeOH, 0-6%] to afford the title compound [745 mg, (94%)]. LC-MS m/z 671 (M+H)⁺.

Example 153 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

The mixture of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (300 mg), 3-[(methyloxy)carbonyl]benzoic acid (180 mg), HBTU (417 mg) and DIPEA (690 μL) in dichloromethane (DCM, 25 mL) was stirred at room temperature overnight. It was concentrated. Separation via CombiFlash (MeOH, DCM) then afforded the desired product (670 mg).

Example 154 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

The mixture of methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (130 mg), LiOH (67 mg), THF (9 mL) and water (3 mL) was stirred at room temperature overnight. It was acidified with HCl to pH˜3. It was extracted with EtOAc and the extracts were combined and dried over Na₂SO₄. Removing the solvent provided the desired product (137 mg).

Example 155 1,1-Dimethylethyl (2S)-4-[(3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate

A mixture of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (69 mg), 1,1-dimethylethyl (2S)-4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate (60 mg), HBTU (68 mg) and diisopropyl ethyl amine (112.5 μL) in DCM (5 mL) and THF (1 mL) were stirred at room temperature overnight. It was concentrated. Separation via CombiFlash then afforded the desired product (33 mg).

Example 156 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (2.0 g, 3.0 mmol) in THF (50 mL) was added MnO₂ (5.2 g, 60.0 mmol). The resulting solution was stirred at room temperature overnight. Then the solution was filtered though Celite. The solvent was removed and 2.0 g of the title compound were collected. Finally, LC-MS was taken to confirm structures. LC-MS m/z 664 (M+H)⁺, 1.86 min (ret time).

Example 157 1,1-Dimethylethyl 4-[(3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-1-piperazinecarboxylate

To a solution of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.072 g, 0.16 mmol) in DMF (1 mL) was added 1,1-dimethylethyl 4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.061 g, 0.16 mmol), (1-hydroxy-1H-benzotriazolato-O)tri-1-pyrrolidinylphosphorus hexafluorophosphate (0.100 g, 0.192 mmol), and TEA (0.045 mL, 0.320 mmol) and this mixture was stirred at room temperature for 16 h. The resultant mixture was purified using a Gilson HPLC (with 0.1% TFA) and concentrated by GeneVav EZ-2 evaporator to afford the title compound. It was used in the next reaction step without further purification. LC-MS m/z 815 (M+H)⁺.

Example 158 1,1-Dimethylethyl 4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-1-piperazinecarboxylate

To a solution of [(3-bromophenyl)methyl]amine hydrochloric salt (0.556 g, 2.5 mmol) in 1,4-dioxane (15 mL) and H₂O (5 mL) was added {3-[(4-{[(1,1-dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]phenyl}boronic acid (1.041 g, 3.25 mmol), Pd(PPh₃)₄ (0.144 g, 0.125 mmol), K₂CO₃ (1.382 g, 10 mmol) and was heated in a microwave at 150° C. for 15 min. The organic layer was separated, dried with a Glas-Col evaporator, then was purified by CombiFlash chromatograph to afford 0.798 g (84%) of the title compound. LC-MS m/z 382 (M+H)⁺.

Example 159 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

To a solution of methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (705 mg, 1.514 mmol) in MeOH (4.5 mL) and H₂O (2.250 mL) was added LiOH (181 mg, 7.57 mmol). This mixture was heated in a microwave at 80° C. for 30 min. The reaction mixture was concentrated, acidified with HCl to ph ˜5, filtered, and washed with MeOH/H₂O (20%) to afford 0.5813 g (85%) of the title compound. LC-MS m/z 452 (M+H)⁺.

Example 160 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

To a solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (1.02 g, 3 mmol) in DCM (30 mL) was added 3-[(methyloxy)carbonyl]benzoic acid (541 mg, 3 mmol), EDC (633 mg, 3.3 mmol), and TEA (1.05 mL, 7.5 mmol). This mixture was stirred at room temperature for 18 h. The reaction mixture was washed with H₂O (20 mL), dried over Na₂SO₄, filtered, concentrated and purified with CombiFlash to afford 0.7052 g (50%) of the title compound. LC-MS m/z 466 (M+H)⁺.

Example 161 1,1-Dimethylethyl 4-[(3′-{[({4-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino) carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-1-piperazinecarboxylate

To a solution of 4-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.072 g, 0.16 mmol) in DMF (1 mL) was added 1,1-dimethylethyl 4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.061 g, 0.160 mmol), HBTU (0.100 g, 0.192 mmol), and TEA (0.045 mL), 0.320 mmol). This mixture was stirred at room temperature for 3 h. The reaction mixture was purified using a Gilson HPLC (with 0.1% TFA) to afford the title compound. LC-MS m/z 815 (M+H)⁺.

Example 162 N-[(3-Bromo-4-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

To a solution of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (500 mg, 1.107 mmol) in DCM (11 mL) was added [(3-bromo-4-fluorophenyl)methyl]amine (266 mg, 1.107 mmol), HBTU (504 mg, 1.329 mmol), and TEA (0.386 mL, 2.77 mmol). This mixture was stirred at room temperature for 18 h before being purified by CombiFlash chromatography to afford 0.6767 g (96%) of the title compound. LC-MS m/z 637 (M+H)⁺.

Example 163 1,1-Dimethylethyl 4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperidinecarboxylate

To a solution of 1,1-dimethylethyl 4-[(3-bromophenyl)methyl]-1-piperidinecarboxylate (191 mg, 0.539 mmol) in DMF (1 mL) was added PdCl₂(dppf) (15.78 mg, 0.022 mmol), bis(pinacolato)diboron (144 mg, 0.566 mmol), and potassium acetate (106 mg, 1.078 mmol). This mixture was placed in a microwave at 100° C. for 1 h. The reaction mixture was diluted with EtOAc (10 mL), washed with H₂O (3×3 mL), brine (3 mL) and the EtOAC dried over Na₂SO₄. The mixture was filtered, concentrated and purified by CombiFlash chromatograph to afford 0.162 g (74%) of the title compound. LC-MS m/z 402 (M+H)⁺.

Example 164 1,1-Dimethylethyl 4-[(3-bromophenyl)methyl]-1-piperidinecarboxylate

To a solution of 1,1-dimethylethyl 4-[(3-bromophenyl)methylidene]-1-piperidinecarboxylate (201 mg, 0.571 mmol) in THF (1 mL) was applied onto H-Cube using 10% Pd/C at a flow rate of 1 mL/min and 1 atm H₂. The mixture was concentrated to afford 0.1909 g (94%) of the title compound. LC-MS m/z 354 (M+H)⁺.

Example 165 1,1-Dimethylethyl 4-[(3-bromophenyl)methylidene]-1-piperidinecarboxylate

To [(3-bromophenyl)methyl](triphenyl)phosphonium bromide (1.13 mg, 2.2 mmol) in DMF (4 mL) was added NaH (52.8 mg, 2.2 mmol) after which the mixture was stirred at RT for 5 min before 1,1-dimethylethyl 4-oxo-1-piperidinecarboxylate (400 mg, 2.0 mmol) was added. This mixture was stirred at RT for 1 h. The resultant mixture was diluted with Et₂O (25 mL), washed with H₂O (12+2×8 mL), brine (8 mL), dried over NaSO₄, and filtered. The filtrate was concentrated and purified with CombiFlash chromatograph to afford the title compound (0.2014 g, 28%). LC-MS m/z 352 (M+H)⁺.

Example 166 [(3-Bromophenyl)methyl](triphenyl)phosphonium Bromide

To 1-bromo-3-(bromomethyl)benzene (2.5 g, 10.0 mmol) in toluene (15 mL) was added triphenylphosphane (2.62 g, 10.0 mmol). This mixture was heated in a microwave oven at 100° C. for 1 h. The mixture was then filtered to afford the title compound (4.55 g, 89%). LC-MS m/z 431 (cationic part)⁺.

Example 167 1-(3-Bromophenyl)-N-methylmethanamine

To bromobenzylamine (0.890 g, 4 mmol) in THF (9 mL) was added NaOH (4.20 mL, 1 N, 4.20 mmol) and the mixture was stirred at room temperature for 5 min where upon Boc₂O (0.975 mL, 4.20 mmol) was added and that mixture was stirred for an additional 30 min. The reaction mixture was diluted with EtOAc (20 mL). The organic layer was separated, washed with brine (5 mL), dried over Na₂SO₄, filtered and concentrated. LAH (12.00 mL, 12.00 mmol) was added to the above crude product and heated in a microwave at 100° C. for 1 h. The reaction was quenched and diluted with Et₂O (˜50 mL) and quenched slowly with Na₂SO₄ (sat.). The organic layer was separated, dried over Na₂SO₄, filtered, and concentrated to afford the title compound (0.472 g, 59%). LC-MS m/z 200 (M+H)⁺.

Example 168 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

To N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.665 g, 1 mmol) in THF (20 mL) was added MnO₂ (0.869 g, 10.00 mmol) and the mixture was stirred at room temperature for 16 h. More MnO₂ (10 eq) was added and stirred at room temperature for 4 h and then again for 22 h and then more MnO₂ (10 eq) was added. This mixture was stirred at room temperature for 21 h and 94 h then filtered and concentrated to afford the title compound (0.6472 g, 98%). LC-MS m/z 663 (M+H)⁺.

Example 169 Methyl 3-[(4-hydroxy-1-piperidinyl)carbonyl]benzoate

To 3-[(methyloxy)carbonyl]benzoic acid (0.901 g, 5 mmol) in DCM (25 mL) was added TEA (0.697 mL, 5.00 mmol), and EtOCOCl (0.480 mL, 5.0 mmol), stirred at 0° C. for 10 min. and then 4-piperidinol (0.506 g, 5.00 mmol) was added. This mixture was then stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (35 mL), washed with HOAc (20 mL, 10%), NaHCO₃ (20 mL, 10%), water (20 mL), dried over Na₂SO₄, filtered and concentrated and purified by CombiFlash chromatograph to afford the title compound (0.696 g, 53%). LC-MS m/z 264 (M+H)⁺.

Example 170 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic Acid

To methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate (3.51 g, 7.54 mmol) in methanol (22.6 mL) and water (11.30 mL), was added LiOH.H₂0 (1.582 g, 37.7 mmol). This was stirred at room temperature for 19 h. MeOH was removed under vacuo. The aqueous layer was acidified with HCl (1 N) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered and concentrated to afford the title compound (2.2082 g, 65%). LC-MS m/z 452 (M+H)⁺.

Example 171 Methyl 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoate

To 3-(methoxycarbonyl)benzoic acid (1.261 g, 7.00 mmol) in DCM (35 mL) was added TEA (2.439 mL, 17.50 mmol), EtOCOCl (0.739 mL, 7.70 mmol) at 0° C. after which 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (2.379 g, 7.00 mmol) was added. This mixture was then stirred for 1 h at room temperature. The reaction mixture was diluted with DCM (35 mL) then washed with HOAc (2×20 mL, 10%), NaHCO₃ (20 mL, 10%), and water (20 mL), concentrated, The residue washed with EtOAc/Hexane (10 mL, 1:1) to afford the title compound (3.5138 g, 94.2%). LC-MS m/z 466 (M+H)⁺.

Example 172 tert-Butyl 4-{[3′-(aminomethyl)biphenyl-3-yl]methyl}piperidine-1-carboxylate

1,1-Dimethylethyl 4-[(3′-cyano-3-biphenylyl)methylidene]-1-piperidinecarboxylate (0.517 g, 1.381 mmol) in methanol (138 mL) was applied on H-Cube hydrogenation apparatus. This resultant mixture was run with Pd(OH)₂ cartridge at 1 mL/min, with 1 atmosphere at 20° C. HCl (1.38 mL, 1 N) was then added. One portion was run one time with a Pd(OH)₂ cartridge at 1 mL/min, 1 atmosphere at 20° C. Another portion was run with 1 run with a Pd(OH)₂ cartridge at 1 mL/min with 50 atmosphere and 20° C. Both portions were combined for 1 run using a Pd(OH)₂ cartridge, under conditions of 1 mL/min with 1 atmosphere at 20° C. The reaction mixture was concentrated to afford the title compound (0.5008 g, 87%). LC-MS m/z 381 (M+H)⁺.

Example 173 1,1-Dimethylethyl 4-[(3′-cyano-3-biphenylyl)methylidene]-1-piperidinecarboxylate

To 1,1-dimethylethyl 4-[(3-bromophenyl)methylidene]-1-piperidinecarboxylate (2.11 g, 5.99 mmol) in 1,4-dioxane (30 mL) and water (10.00 mL) was added m-NC(C₆H₄)B(OH)₂ (1.056 g, 7.19 mmol), Pd(Ph₃P)₄ 0.277 g, 0.240 mmol), and K₂CO₃ (2.483 g, 17.97 mmol). The resultant mixture was split into two equal portions and each portion was heated in a microwave at 130° C. for 15 min. The reaction mixture was concentrated by Glas-Col evaporator and purified with CombiFlash chromatograph to afford the title compound (1.93 g, 86%). LC-MS m/z 749 (2M+H)⁺.

Example 174 [1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]acetic Acid

To [1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]acetonitrile (0.667 g, 2.128 mmol) in Ethanol (10 mL) was added a KOH 40% aqueous solution (10 mL). This mixture was heated in a microwave at 100° C. for 1 h, then heated again in the microwave at 100° C. for 10 h. This was repeated with microwave heating at 120° C. for 1 h and yet another time it was heated in the microwave at 120° C. for and additional 5 h. Next, EtOH was removed under vacuum and the residual material was acidified to pH˜5 and extracted with DCM/i-PrOH (3/1, 2×30 mL). This mixture was concentrated and purified using a Gilson HPLC (with TFA) to afford the title compound (0.3171 g, 45%). LC-MS m/z 333 (M+H)⁺.

Example 175 N-[(3-Bromo-2-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

A mixture of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (210 mg, 0.466 mmol), [(3-bromo-2-fluorophenyl)methyl]amine hydrochloride (123 mg, 0.513 mmol), HBTU (194 mg, 0.513 mmol) and diisopropyl ethyl amine (0.5 mL) in DCM (10 mL) was stirred at room temperature for about 5 h. It was concentrated, mixed with EtOAc (25 mL), washed with aqueous NaHCO₃ (25 mL) and then NH₄Cl (25 mL). The organic layer was collected, dried and concentrated to afford the desired product as a crude mixture (330 mg).

Example 176 3-{[({3′-[(4-{[(1,1-Dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)amino]sulfonyl}benzoic Acid

To a solution of 3-(chlorosulfonyl)benzoic acid (0.0331 g, 0.15 mmol) in DCM (1.5 mL) was added 1,1-dimethylethyl 4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.060 g, 0.15 mmol), and Et₃N (0.0211 mL, 0.15 mmol). The resulting mixture was stirred at room temperature for 1 h before it was concentrated and purified using a Gilson HPLC to afford the title compound 0.0374 g (43%). LC-MS m/z 584 (M+H)⁺.

Example 177 3-{[[(3-Bromophenyl)methyl](methyl)amino]sulfonyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}benzamide

To 3-{[[(3-bromophenyl)methyl](methyl)amino]sulfonyl}benzoic acid (150 mg, 0.390 mmol) in DCM 7.8 mL) was added 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (133 mg, 0.390 mmol), HBTU (163 mg, 0.429 mmol), and TEA (0.163 mL, 1.171 mmol). This mixture was stirred at room temperature for 2 h. The reaction mixture was washed with H₂O (2×3 mL), dried over Na₂SO₄, filtered, and concentrated. The crude product was further purified by CombiFlash flash chromatograph to afford the title compound (0.2453 mg, 94%). LC-MS m/z 669 (M+H)⁺.

Example 178 3-{[[(3-Bromophenyl)methyl](methyl)amino]sulfonyl}benzoic Acid

1-(3-Bromophenyl)-N-methylmethanamine (0.100 g, 0.5 mmol) in DCM (5 mL) was added TEA (0.139 mL, 1.000 mmol) and 3-(chlorosulfonyl)benzoic acid (0.110 g, 0.500 mmol). The resulting mixture was stirred at room temperature for 0.5 h. The reaction was quenched with HCl (1 N), concentrated and redissolved in EtOAc (10 mL) which was washed with H₂O (2×3 mL), brine (3 mL), then dried with Na₂SO₄. It was then filtered and concentrated to afford the title compound (0.1505 g, 78%). LC-MS m/z 384 (M+H)⁺.

Example 179 1,1-Dimethylethyl 4-[(3′-cyano-3-biphenylyl)methylidene]-1-piperidinecarboxylate

To a solution of 1,1-dimethylethyl 4-[(3-bromophenyl)methylidene]-1-piperidinecarboxylate (2.07 g, 5.88 mmol) in p-dioxane (30 mL) and H₂O (10 mL) was added (3-cyanophenyl)boronic acid (1.036 g, 7.05 mmol), Pd(PPh₃) (0.272 g, 0.235 mmol), and K₂CO₃ (2.436 g, 17.63 mmol) and then the reaction was heated in a microwave at 130° C. for 15 minutes. The organic layer of the reaction mixture was separated, concentrated and purified by Combiflash chromatograph to afford 1.88 g (85%) of the title compound. LC-MS m/z 749 (2M+H)⁺.

Example 180 1,1-Dimethylethyl 4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-1-piperidinecarboxylate Hydrochloride

To a solution of 1,1-dimethylethyl 4-[(3′-cyano-3-biphenylyl)methylidene]-1-piperidinecarboxylate (1.87 g, 4.99 mmol) in MeOH (499 mL) was added HCl (4.99 mL, 1N, 4.99 mmol) then was applied to H-cube hydrogenation apparatus until the reaction was complete per monitoring with LC-MS. The reaction mixture was concentrated to afford 1.40 g (67%) of the title compound. LC-MS m/z 381 (M+H)⁺.

Example 181 1-{3′-[(1-Methyl-4-piperidinyl)methyl]-3-biphenylyl}methanamine

To 1,1-dimethylethyl 4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-1-piperidinecarboxylate hydrochloride (1.23 g, 2.95 mmol) was added LiAlH₄ (5.90 mL, 1N in THF, 5.90 mmol) and the reaction was stirred at room temperature until the was complete per monitoring with LC-MS. The reaction mixture was diluted with Et₂O (50 mL) and quenched with Na₂CO₃ (saturated aqueous), filtered, concentrated and purified with Combiflash chromatograph to afford 0.59 g (68%) of the title compound. LC-MS m/z 295 (M+H)⁺.

Example 182 3-{[({3′-[(1-Methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)amino]sulfonyl}benzoic Acid

To a solution of 1-{3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methanamine (0.0589 g, 0.2 mmol) in DCM (2 mL) was added TEA (0.0606 mL, 0.4 mmol) and then 3-(chlorosulfonyl)benzoic acid (0.0441 g, 0.2 mmol) whereupon the reaction was stirred at room temperature for 17 h. The reaction mixture was concentrated on a Glas-Col evaporator, purified using a Gilson HPLC (with 0.1% TFA), and concentrated to afford 0.0861 g (57%) of the title compound. LC-MS m/z 479 (M+H)⁺.

Example 183 3-[(3-Bromophenyl)methylidene]-8-methyl-8-azabicyclo[3.2.1]octane

To a solution of diethyl [(3-bromophenyl)methyl]phosphonate (3.07 g, 10 mmol) in THF (20 mL) was added 8-methyl-8-azabicyclo[3.2.1]octan-3-one (1.67 g, 12 mmol) then KOBu-t (1.234 g, 11 mmol). The reaction mixture was stirred at room temperature for 5 h before it was quenched with NH₄Cl (5 mL, saturated aqueous). The organic layer of the reaction mixture was separated and the aqueous layer was extracted with DCM (5 mL). The combined organic layers were loaded onto a Combiflash Redisep silica gel column (40 g) and purified by Combiflash chromatograph to afford 2.48 g (85%) of the title compound. LC-MS m/z 292 (M+H)⁺.

Example 184 3′-[(8-Methyl-8-azabicyclo[3.2.1]oct-3-ylidene)methyl]-3-biphenylcarbonitrile

To a solution of 3-[(3-bromophenyl)methylidene]-8-methyl-8-azabicyclo[3.2.1]octane (1.17 g, 4 mmol) in p-dioxane (15 mL) and H₂O (5 mL) was added (3-cyanophenyl) boronic acid (0.705 g, 4.80 mmol), Pd(PPh₃) (0.185 g, 0.16 mmol), and K₂CO₃ (1.658 g, 12 mmol) and then the mixture was heated in a microwave oven at 130° C. for 20 min. The organic layer of the reaction mixture was separated, concentrated and purified by Combiflash chromatograph to afford 1.22 g (97%) of the title compound. LC-MS m/z 315 (M+H)⁺.

Example 185 1-{3′-[(8-Methyl-8-azabicyclo[3.2.1]oct-3-yl)methyl]-3-biphenylyl}methanaminedihydrochloride

To a solution of 3′-[(8-methyl-8-azabicyclo[3.2.1]oct-3-ylidene)methyl]-3-biphenylcarbonitrile (1.21 g, 3.85 mmol) in MeOH (192 mL) was added HCl (3.85 mL, 1N, 3.85 mmol). This mixture was then applied to an H-cube hydrogenation apparatus using 20% Pd(OH)₂ cartridge at a flow rate of 1 mL/min. and H₂ (1 atm) at 20° C. until reaction complete by monitoring with LC-MS. The reaction mixture was concentrated to afford 1.31 g (87%) of the title compound. LC-MS m/z 321 (M+H)⁺.

Example 186 1,1-Dimethylethyl (2S)-4-{[3-(4-cyano-2-pyridinyl)phenyl]methyl}-2-methyl-1-piperazinecarboxylate

To 2-chloro-4-pyridinecarbonitrile (0.416 g, 3 mmol) was added 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (1.499 g, 3.60 mmol) followed by K₂CO₃ (1.244 g, 9.00 mmol) and Pd(Ph₃P)₄ (0.139 g, 0.120 mmol) in 1,4-dioxane (15.00 mL) and water (5.00 mL). The mixture was heated in a microwave oven for 30 min at 140° C. The organic layer was collected and the aqueous layer was extracted once with EtOAc (3 mL). The combined organic layers were filtered, concentrated on Glas-Col, redissolved in hexane/DCM (3:1, 4 mL), loaded onto Redisep silica gel column (12 g) and purified with Combiflash chromatography. Fractions were collected and the solvent was removed under reduced pressure to afford 1.10 g (94%) of the title compound. LC-MS m/z 393 (M+H)⁺.

Example 187 1,1-Dimethylethyl (2S)-4-({3-[4-(aminomethyl)-2-pyridinyl]phenyl}methyl)-2-methyl-1-piperazinecarboxylate

To 1,1-dimethylethyl (2S)-4-{[3-(4-cyano-2-pyridinyl)phenyl]methyl}-2-methyl-1-piperazinecarboxylate (1.20 g, 3.06 mmol) was added methanol (306 mL) to give a 0.01 molar yellow solution. Then the solution was applied to the H-Cube hydrogenation apparatus: 1st run, 10% Pd/C cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 2nd run, 10% Pd/C cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 3rd run, 10% Pd/C cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction completed). The mixture was concentrated, redissolved in hexane/DCM (3:1, 5 mL), loaded onto Redisep silica gel column (40 g) and purified with Combiflash chromatograph to afford 937 mg (77%) of the title compound. LC-MS m/z 397 (M+H)⁺.

Example 188 1,1-Dimethylethyl 4-{[3-(4-cyano-2-pyridinyl)phenyl]methyl}-1-piperidinecarboxylate

To 2-chloro-4-pyridinecarbonitrile (416 mg, 3 mmol) was added {3-[(1-{[(1,1-dimethylethyl)oxy]carbonyl}-4-piperidinyl)methyl]phenyl}boronic acid (958 mg, 3.00 mmol) followed by K₂CO₃ (1,244 mg, 9.00 mmol) and Pd(Ph₃P)₄ (139 mg, 0.120 mmol) in 1,4-dioxane (15 mL) and water (5 mL). The mixture was heated in a microwave oven for 30 min at 140° C. The organic layer was collected and the aqueous layer was extracted with EtOAc (3 mL). The combined organic layers were filtered, concentrated on Glas-Col, redissolved in hexane/DCM (3:1, 4 mL), loaded onto Redisep silica gel column (40 g) and purified with Combiflash chromatograph to afford 640 mg (57%) of the title compound. LC-MS m/z 378 (M+H)⁺.

Example 189 1,1-Dimethylethyl 4-({3-[4-(aminomethyl)-2-pyridinyl]phenyl}methyl)-1-piperidinecarboxylate

To 1,1-dimethylethyl 4-{[3-(4-cyano-2-pyridinyl)phenyl]methyl}-1-piperidinecarboxylate (640 mg, 1.695 mmol) was added methanol (1,695 μl) to give a 0.01 molar solution. Then the solution was applied to the H-Cube hydrogenation apparatus: 1st run, 20% Pd(OH) cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 2nd run, 20% Pd(OH) cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 3rd run, 20% Pd(OH) cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 4th run, 20% Pd(OH) cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 5th run, 20% Pd(OH) cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 6th run, 20% Pd(OH) cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction not completed); 7th run, 20% Pd(OH) cartridge, H₂ (1 atm), 1 mL/min, 20° C. (reaction completed). The mixture was concentrated, redissolved in hexane/DCM (3:1, 5 mL), loaded onto Redisep gel column (40 g) and purified with Combiflash chromatograph to afford 413 mg (64%) of the title compound. LC-MS m/z 382 (M+H)⁺.

EXAMPLES Compounds of Formula (I) Example 190 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(4-morpholinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.5 g, 0.75 mmol) in dichloromethane (10 mL) was added thionyl chloride (1 mL) and the mixture was heated at reflux for 2 h and then concentrated to dryness. To 170 mg of the residual material in dichloromethane (5 mL) was added morpholine (0.022 mL, 0.2 mmol) and diisopropylethylamine (0.087 mL. 0.54 mmol) and the resulting solution was stirred at ambient temperature for 16 h. The mixture was then concentrated to dryness and the product purified by mass-directed automated preparative HPLC. Product-containing fractions were passed though a 2 g SCX solid-phase extraction cartridge and the product eluted off with a 10% solution of ammonia in methanol, product containing fractions were concentrated to dryness, dissolved in methanol (10 mL) and added to a 2 g SCX solid-phase extraction cartridge and the product eluted off with a 10% solution of pyridine in methanol. The product-containing fractions were concentrated to dryness and further purified by mass-directed automated preparative HPLC to afford 7 mg of the title compound. LC-MS m/z 734 (M+H)⁺, 0.83 min (ret time).

Example 191 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.5 g, 0.75 mmol) in dichloromethane (10 mL) was added thionyl chloride (1 mL) and the mixture was heated at reflux for 2 h and then concentrated to dryness. To 170 mg of the residual material in dichloromethane (5 mL) was added 1-(tert-butyloxycarbonyl)piperazine (0.045 mL, 0.25 mmol) and diisopropylethylamine (0.087 mL. 0.54 mmol) and the resulting solution was stirred at ambient temperature for 16 h, at 50° C. for 2 h, 65° C. for 3 h and then 75° C. for 4 h. The cooled mixture was treated with a solution of hydrochloric acid in 1,4-dioxane (4M, 5 mL) and stirred at ambient temperature for 16 h. The mixture was then concentrated to dryness and the product purified by mass-directed automated preparative HPLC. Product-containing fractions were passed though a 2 g SCX solid-phase extraction cartridge and the product eluted off with a 10% solution of ammonia in methanol, product containing fractions were concentrated to dryness to afford 29 mg of the title compound. LC-MS m/z 733 (M+H)⁺, 0.82 min (ret time).

Example 192 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (0.033 g, 0.05 mmol), and piperazine (0.043 g, 0.5 mmol) were combined in CH₂Cl₂ (2 mL) and stirred for 30 min. Sodium triacetoxyborohydride (0.0318 g, 0.15 mmol) was added and the mixture stirred for 30 h. Aqueous workup was followed using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 1 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 729.4 (M+H)⁺, 1.31 min (ret time); mp 135-138° C. Analytical HPLC shows 98.4% purity, (ret time 10.019 min).

Example 193 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (0.033 g, 0.05 mmol) and 1-methylpiperazine (0.025 g, 0.25 mmol) were combined in CH₂Cl₂ (2 mL) and stirred for 30 min. Sodium triacetoxyborohydride (0.053 g, 0.25 mmol) was added and the mixture stirred overnight. Aqueous workup followed using a Gilson HPLC with an ammonium hydroxide system gave the title compound as a white solid. LC-MS m/z 743.2 (M+H)⁺, 1.42 min (ret time); mp 121-124° C.

Example 194 N-{[3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-6-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.0337 g, 0.05 mmol), 1,1-dimethylethyl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (0.0119 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL). Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the protected product. This was treated with 10% TFA in CH₂Cl₂. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 1 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 757.2 (M+H)⁺, 1.32 min (ret time); mp 129-131° C. Analytical HPLC shows 100% purity, (ret time 9.790 min).

Example 195 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.0337 g, 0.05 mmol), 1,1-dimethylethyl hexahydro-1H-1,4-diazepine-1-carboxylate (0.012 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL), and the mixture stirred for thirty min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the protected product. This was treated with 10% TFA in CH₂Cl₂. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 4× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the tile compound as a tan solid. LC-MS m/z 759.2 (M+H)⁺, 1.32 min (ret time); mp 143-147° C. Analytical HPLC shows 98.4% purity, (ret time 9.755 min).

Example 196 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-6-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (0.033 g, 0.05 mmol), 1,1-dimethylethylhexahydro-1H-1,4-diazepine-1-carboxylate (0.012 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL), and the mixture stirred for thirty min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the protected product. This was treated with 10% TFA in CH₂Cl₂ for 3 h. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a light-yellow solid. LC-MS m/z 743.2 (M+H)⁺, 1.26 min (ret time); mp 118-120° C. Analytical HPLC shows 99.4% purity, (ret time 9.965 min).

Example 197 N-({3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-6-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

Process (A) N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide, (0.033 g, 0.05 mmol), 1,1-dimethylethyl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (0.0119 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL), and the mixture stirred for thirty min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the protected product. This was treated with 10% TFA in CH₂Cl₂ for 3 h. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 741.2 (M+H)⁺, 1.29 min (ret time); mp 149-152° C. Analytical HPLC shows 100% purity, (ret time 9.997 min).

Process (B) In an alternate process for preparing the compound of Example 184, N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (57 mg, 0.087 mmol) and 1,1-dimethylethyl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (36.1 mg, 0.173 mmol) were dissolved in 1,2-Dichloroethane (DCE) (1 mL) in a 1 dram vial and the acetic acid (5.45 μL, 0.095 mmol) was added. The mixture was stirred for 30 minutes and then the MP-Triacetoxyborohydride (186 mg, 0.433 mmol) was added. The mixture was stirred overnight and then filtered through glass fiber filter paper and washed 2× with 0.3 mL of 1,2-dichloroethane. Trifluoroacetic acid (0.18 mL, 2.336 mmol) was added and the mixture stirred at room temperature overnight. The reaction mixture was concentrated and taken up in 1.4 mL of DMSO, and purified on a Gilson HPLC eluting at 20 mL/min with a linear gradient running from 10% CH₃CN/H₂O (0.1% TFA) to 90% CH₃CN/H₂O (0.1% TFA). The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated. The organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-6-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a white solid (43.3 mg, 67.5%). LC-MS m/z 742 (M+H)⁺; mp 138-140° C.; ¹H NMR (400 MHz, CD₃OD) δ 1.28-1.35 (m, 3 H) 1.41 (t, J=7.15 Hz, 3 H) 1.58 (d, J=10.54 Hz, 1 H) 1.68-1.76 (m, 2 H) 1.86-1.92 (m, 1 H) 2.00-2.08 (m, 2 H) 2.20 (s, 3 H) 2.55 (dd, J=10.29, 1.25 Hz, 1 H) 2.77 (dd, J=10.67, 2.38 Hz, 1 H) 2.85 (dd, J=10.29, 2.51 Hz, 1 H) 3.01 (q, J=7.53 Hz, 2 H) 3.21 (dd, J=10.67, 1.13 Hz, 1 H) 3.43 (s, 1 H) 3.58-3.65 (m, 3 H) 3.76 (q, J=13.05 Hz, 2 H) 3.94-4.02 (m, 2 H) 4.12-4.20 (m, 1 H) 4.42 (q, J=7.28 Hz, 2 H) 4.56 (s, 2 H) 4.70 (s, 2 H) 7.16-7.19 (m, 2 H) 7.20-7.25 (m, 2 H) 7.28-7.35 (m, 3 H) 7.55 (t, J=7.78 Hz, 1 H) 7.98 (dd, J=8.28, 2.26 Hz, 3 H) 8.29 (t, J=1.51 Hz, 1 H).

Example 198 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(2R,5S)-2,5-dimethyl-1-piperazinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide. (0.041 g, 0.06 mmol), trans-2,5-dimethylpiperazine (0.034 g, 0.3 mmol), and N,N-diisopropylethylamine (52 μL, 0.3 mmol) were combined in CH₂Cl₂ (2 mL) and stirred under argon at to 40° C. for 48 h. As the reaction had not gone to completion the mixture was then microwaved at 75° C. for 20 min, and then microwaved at 90° C. for 40 min. Additional trans-2,5-dimethylpiperazine (0.034 g, 0.3 mmol) and diisopropylethylamine (52 μL, 0.3 mmol) were added, and the mixture microwaved at 100° C. for 2 h. Additional trans-2,5-dimethylpiperazine (0.034 g, 0.3 mmol) and diisopropylethylamine (52 μL, 0.3 mmol) were added, and the mixture microwaved at 100° C. for 1 h. Little or no starting material remained. The reaction mixture was partitioned between EtOAc and water. The organic phase was washed 3× with water to remove the excess trans-2,5-dimethylpiperazine. The EtOAc phase was concentrated to a small volume, and applied to a 2 g SCX column. This was washed with EtOAc, and then with MeOH, and finally with 2 M NH₃ in MeOH. The Fractions containing the impure product was purified using a Gilson HPLC to give the TFA salt of the desired product. This was passed though a quaternary ammonium hydroxide column to give after evaporation the title compound as a tan solid. LC-MS m/z 760.8 (M+H)⁺, 1.42 min (ret time); mp 187-190° C. (dec). Analytical HPLC shows 98.6% purity, (ret time 9.977 min).

Example 199 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.0455 g, 0.0667 mmol) and piperidine (66 μL, 0.667 mmol) were combined in THF (2.5 mL) and microwaved at 80° C. for 1 h. The reaction mixture was concentrated and taken up in EtOAc, washed with water 2×, brine 1×, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. This was purified using a Gilson HPLC followed by passing though 2 quaternary ammonium hydroxide cartridges to give the free base, the title compound, as a tan solid. LC-MS m/z 731.8 (M+H)⁺, 1.60 min (ret time); mp 128-131° C. Analytical HPLC shows 99.1% purity, (ret time 11.149 min).

Example 200 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-AN-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.025 g, 0.036 mmol) and 1-methylpiperazine (0.018 g, 0.18 mmol) were combined in THF (1 mL) and microwaved at 80° C. for 1 h, and then at 90° C. for 1 h. The mixture was purified using a Gilson HPLC followed by aqueous workup and then passing product though a quaternary ammonium hydroxide cartridge to give the free base, the title compound, as a white solid. LC-MS m/z 746.8 (M+H)⁺, 1.41 min (ret time); mp 169-173° C. Analytical HPLC shows 96.4% purity, (ret time 10.018 min).

Example 201 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(3-oxo-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.025 g, 0.036 mmol) and 2-piperazinone (0.018 g, 0.18 mmol) were combined in THF (1 mL) and microwaved at 80° C. for 1 h, then at 90° C. for 1 h, then 100° C. for 2.5 h, 120° C. for 1 h, and then 130° C. for 1 h. The mixture was purified using a Gilson HPLC followed by passing product though a quaternary ammonium hydroxide cartridge to give the free base, the title compound, as a white solid. LC-MS m/z 746.8 (M+H)⁺, 1.40 min (ret time); mp 147-150° C. Analytical HPLC shows 100% purity, (ret time 10.329 min).

Example 202 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(4-ethyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol) and 1-ethylpiperazine (0.0285 g, 0.25 mmol) were combined in THF (1 mL) and microwaved at 150° C. for 1 h. The mixture was purified using a Gilson HPLC with a TFA system followed by passing the product though a quaternary ammonium hydroxide cartridge to give the free base, the title compound, as a tan solid. LC-MS m/z 760.8 (M+H)⁺, 1.55 min (ret time); mp 155-158° C. Analytical HPLC shows 95.7% purity, (ret time 10.098 min).

Example 203 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(1-methylethyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol) and 1-(1-methylethyl)piperazine (0.0325 g, 0.25 mmol) were combined in THF (1 mL) and microwaved at 150° C. for 1 h. The mixture was purified using a Gilson HPLC with a TFA system followed by passing product though a quaternary ammonium hydroxide cartridge to give the free base, the title compound, as a white solid. LC-MS m/z 774.8 (M+H)⁺, 1.46 min (ret time); mp 165-169° C. Analytical HPLC shows 100% purity, (ret time 10.233 min).

Example 204 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol) and homopiperazine (0.025 g, 0.25 mmol) were combined in THF (1 mL) and microwaved at 150° C. for 1 h. The mixture was purified using a Gilson HPLC with a TFA system was followed by passing product though a quaternary ammonium hydroxide cartridge to give the free base, the title compound, as a white solid. LC-MS m/z 746.8 (M+H)⁺, 1.38 min (ret time); mp 145-148° C. Analytical HPLC shows 99.6% purity, (ret time 9.774 min).

Example 205 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

Process (A):

205(a) 3-{[({3′-[((3S)-4-{[(1,1-Dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)amino]carbonyl}benzoic acid (1150 mg, 2.0 mmol) was dissolved in dichloromethane (DCM) (100 mL), and then HBTU (830 mg, 2.2 mmol), 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (620 mg, 2.0 mmol), and DIPEA (1.4 mL, 8.0 mmol) were added. The mixture was allowed to stir under argon at room temperature overnight. Solvents were concentrated and then purified on a 40 g Redi-Sep cartridge and eluting with 0.5% MeOH in DCM. Collected clean fractions were combined solvent concentrated to give 1,1-dimethylethyl (2S)-4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate (1.15 g, 66.5%) as a white solid.

205(b) The piperazinecarboxylate intermediate was dissolved in DCM (10 mL), TFA (1.4 mL) was added and mixture was stirred at RT overnight. Solvent was concentrated and the residue was cooled in an ice bath and basified with 15% NH₄OH. It was extracted with ethyl acetate (3×150 mL). The combined organic layers were dried over Na₂SO₄, filtered and then concentrated to give a crude residue that was then purified on a 40 g Redi-Sep cartridge, eluting with 0 to 100% MeOH (10% in DCM) in DCM. Clean fractions were combined and solvent concentrated off to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (780 mg) as a white foamy solid.

Process (B) The benzenedicarboxamide was determined to contain residual TFA based on ¹⁹F NMR. So a further step for the isolation and purification of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide, the compound was undertaken. Material was taken up in EtOAc (100 mL) and washed 3× with 0.5 N NaOH, 1× with brine, and dried over anhydrous Na₂SO₄, filtered, and solvent concentrated to give the title compound. ¹⁹F NMR indicated that all TFA had been removed from the sample. LC-MS m/z 746.8 (M+H)⁺, 1.42 min (ret time); mp 100-113° C. Analytical HPLC shows 99.6% purity, (ret time 9.936 min).

Example 206 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[[2-(dimethylamino)-ethyl](methyl)amino]methyl}-6-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[3′-(Chloromethyl)-6-fluoro-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol) and N,N,N′-trimethyl-1,2-ethanediamine (0.025 g, 0.25 mmol) were combined in THF (1 mL) and microwaved at 150° C. for 1 h. Purification on a Gilson HPLC using a TFA system was followed by passing the product though a quaternary ammonium hydroxide cartridge to give the free base, the title compound, as a white solid. LC-MS m/z 748.8 (M+H)⁺, 1.43 min (ret time); mp 126-129° C. Analytical HPLC shows 99.1% purity, (ret time 9.788 min).

Example 207 N-({3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

1,1-Dimethylethyl (1S,4S)-5-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}-carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (0.0488 g, 0.058 mmol) was dissolved in CH₂Cl₂ (9 mL) and treated with trifluoroacetic acid (1 mL) at 0° C. and then allowed to warm to room temperature. This mixture was stirred for a total of 2 h. The solvents were pumped off, and the material was passed though 2 quaternary ammonium hydroxide cartridges, but ¹⁹F NMR still indicated residual TFA. The product was taken up in EtOAc and washed 3× with 0.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a tan solid. LC-MS m/z 746 (M+H)⁺, 1.30 min (ret time); mp 129-133° C. Analytical HPLC shows 96.8% purity, (ret time 9.820 min).

Example 208 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

Process (A) N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.0337 g, 0.05 mmol), 1-methylpiperazine (0.025 g, 0.25 mmol) were combined in CH₂Cl₂ (2 mL) and stirred for 15 min. Sodium triacetoxy-borohydride (0.053 g, 0.25 mmol) was added and the mixture stirred for 18 h. Aqueous workup followed using a Gilson HPLC with an ammonium hydroxide system gave the title compound as a white solid. LC-MS m/z 759.2 (M+H)⁺, 1.45 min (ret time); mp 117-119° C. Analytical HPLC shows 98.2% purity, (ret time 9.921 min).

Process (B) In an alternate process for the preparation of the title compound N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (34 mg, 0.05 mmol), 1-methylpiperazine (6 mg, 0.06 mmol), MP-Triacetoxyborohydride (43 mg, 0.1 mmol), and acetic acid (3.6 mg, 0.06 mmol) were combined in dimethyl sulfoxide (DMSO) (0.5 ml) in a 1 dram vial and stirred at room temperature overnight. The reaction did not go to completion. Additional 1-methylpiperazine (6 mg, 0.06 mmol) and MP-Triacetoxyborohydride (86 mg, 0.2 mmol) were added to the reaction mixture and stirred for another 2 h. The mixture was filtered through glass fiber filter paper and washed 2× with 0.3 mL of DMSO. The combined filtrates were purified on a Gilson HPLC with the TFA system. The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated and the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-({6-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide as a white solid, mp 120-123° C.

Example 209 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.0337 g, 0.05 mmol), piperazine (0.043 g, 0.5 mmol) were combined in CH₂Cl₂ (2 mL) and stirred for 15 min. Sodium triacetoxyborohydride (0.053 g, 0.25 mmol) was added and the mixture stirred for 18 h. Aqueous workup followed using a Gilson HPLC with a TFA system followed by conversion to the free base by passing though a quaternary ammonium hydroxide cartridge. TFA was still present in ¹⁹F NMR. The compound was taken up in EtOAc and washed 3× with 1 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered and concentrated to give the title compound as a white solid. LC-MS m/z 745.2 (M+H)⁺, 1.52 min (ret time); mp 138-140° C. Analytical HPLC shows 100% purity, (ret time 8.368 min).

Example 210 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.0337 g, 0.05 mmol), (2R,6S)-2,6-dimethylpiperazine (0.00685 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL). Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 773.2 (M+H)⁺, 1.27 min (ret time); mp 127-129° C. Analytical HPLC shows 100% purity, (ret time 9.929 min).

Example 211 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (0.033 g, 0.05 mmol), (2R,6S)-2,6-dimethylpiperazine (0.00685 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL). Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 757.2 (M+H)⁺, 1.37 min (ret time); mp 109-112° C. Analytical HPLC shows 99.8% purity, (ret time 10.205 min).

Example 212 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-methyl-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-[(3-Bromo-4-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.032 g, 0.05 mmol), 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (0.0316 g, 0.076 mmol), potassium carbonate (0.021 g, 0.15 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.0029 g, 0.0025 mmol) were combined in dioxane (1.5 mL) and water (0.5 mL). The mixture was microwaved at 150° C. for 30 min. Then the reaction mixture was diluted with EtOAc and washed with H₂O 3×. The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified via CombiFlash on a 12 g column eluted with 0-10% MeOH/CH₂Cl₂ to give 1,1-dimethylethyl (2S)-4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)amino]methyl}-2′-methyl-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate. This was taken up in 10% TFA/CH₂Cl₂ (2 mL) and stirred at room temperature for 3 h. The solvents were pumped off, and the crude product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 743.2 (M+H)⁺, 1.40 min (ret time); mp 115-118° C. Analytical HPLC shows 100% purity, (ret time 10.211 min).

Example 213 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[3-Bromo-4-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.033 g, 0.05 mmol), 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (0.0316 g, 0.076 mmol), potassium carbonate (0.021 g, 0.15 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.0029 g, 0.0025 mmol) were combined in dioxane (1.5 mL) and water (0.5 mL). The mixture was microwaved at 150° C. for 30 min. The reaction mixture was then diluted with EtOAc and washed with H₂O. The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated. This was taken up in 10% TFA/CH₂Cl₂ and stirred at room temperature for 3 h. The solvents were pumped off and the crude product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed with 2.5 N NaOH, with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound a white solid. LC-MS m/z 759.2 (M+H)⁺, 1.35 min (ret time); mp 110-112° C. Analytical HPLC shows 100% purity, (ret time 9.95 min).

Example 214 N-[(6-Chloro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(3-Bromo-4-chlorophenyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.0327 g, 0.05 mmol) 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (0.0208 g, 0.05 mmol), potassium carbonate (0.021 g, 0.15 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.0029 g, 0.0025 mmol) were combined in dioxane (1.5 mL) and water (0.5 mL). The mixture was microwaved at 100° C. for 15 min. Then additional 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (0.0042 g, 0.01 mmol) was added and the mixture microwaved at 100° C. for 15 min. The reaction mixture was diluted with EtOAc and washed with H₂O. The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated. This was taken up in 10% TFA/CH₂Cl₂ (2 mL) and stirred at room temperature for 3 h. The solvents were pumped off and the crude product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed with 3× with 2.5 N NaOH, then with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 763.1 (M+H)⁺, 1.35 min (ret time); mp 117-120° C. Analytical HPLC shows 100% purity, (ret time 10.254 min).

Example 215 N-[(6-Chloro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

In an alternate process for the preparation of the title compound, N-[(3-bromo-4-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (70 mg, 0.107 mmol), 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (44.6 mg, 0.107 mmol), potassium carbonate (44.4 mg, 0.321 mmol), and Pd(Ph₃P)₄ (6.18 mg, 5.35 μmol) were added to a 0.5-2 mL Biotage microwave vial in 1,4-dioxane (3 mL) and water (1.00 mL). The vial was capped and the mixture was heated under microwave at normal power in the Emrys Optimizer at 100° C. for 15 minutes. The crude product was partitioned between EtOAc (30 mL) and water (10 mL). The phases were separated; the organic washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated to give the crude intermediate. It was then taken up in dichloromethane (DCM) (3.60 mL), and treated with TFA (0.4 mL, 5.19 mmol). The mixture was stirred under argon overnight. Solvents were pumped off and the residue taken up in DMSO (1.1 mL) and purified on a Gilson HPLC eluting at 20 mL/min with a linear gradient running from 10% CH₃CN/H₂O (0.1% TFA) to 90% CH₃CN/H₂O (0.1% TFA). The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking up the residue in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated, the organic phase was washed with 2.5 N NaOH (3×5 mL), and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄ filtered and concentrated to give N-[(6-chloro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (62.3 mg, 76% yield) as a white solid. LC-MS m/z 763 M⁺; mp 116-118° C.

Example 216 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-methyl-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-6-methyl-3-biphenylyl)methyl]-1,3-benzene-dicarboxamide (0.033 g, 0.05 mmol), 1-methylhexahydro-1H-1,4-diazepine (0.007 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL). Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated and the residue taken up in EtOAc. It was washed with 1N NaOH and dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 757.2 (M+H)⁺, 1.38 min (ret time); mp 107-109° C. Analytical HPLC shows 95.3% purity, (ret time 10.035 min).

Example 217 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.0337 g, 0.05 mmol), 1-methylhexahydro-1H-1,4-diazepine (0.007 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in CH₂Cl₂ (2 mL). Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 773.2 (M+H)⁺, 1.32 min (ret time); mp 116-119° C. Analytical HPLC shows 92.8% purity, (ret time 9.808 min).

Example 218 N-({6-Chloro-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol), 1-methylhexahydro-1H-1,4-diazepine (0.0069 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in 1,2-dichloroethane (2 mL). The reaction mixture was stirred for 30 min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 777.1 (M+H)⁺, 1.39 min (ret time); mp 105-107° C. Analytical HPLC shows 95.3% purity, (ret time 10.044 min).

Example 219 N-({6-Chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

Process (A) N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol), 1-methylpiperazine (0.006 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in 1,2-dichloroethane (2 mL). The reaction mixture was stirred for 30 min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 763.1 (M+H)⁺, 1.60 min (ret time); mp 112-114° C. Analytical HPLC shows 100% purity, (ret time 10.367 min).

Process (B) In an alternate process for the preparation of N-({6-chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide, N-[(6-chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (85 mg, 0.125 mmol), 1-methylpiperazine (25.04 mg, 0.250 mmol), MP-Triacetoxyborohydride (268 mg, 0.625 mmol), and acetic acid (7.87 μl, 0.138 mmol) were combined in dimethyl sulfoxide (DMSO) (1.25 mL) in a 1 dram vial and stirred at room temperature overnight. The mixture was filtered through glass fiber filter paper and washed 2× with 0.3 mL of DMSO. The combined filtrates were purified on a Gilson HPLC with the TFA system. The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated and the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-({6-chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (66.1 mg, 69.3%) as a white solid, LC-MS m/z 763 M⁺; mp 112-114° C.

Example 220 N-[(6-Chloro-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol), (2R,6S)-2,6-dimethylpiperazine (0.0069 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in 1,2-dichloroethane (2 mL). The reaction mixture was stirred for 30 min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the crude product. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 777.1 (M+H)⁺, 1.45 min (ret time); mp 121-124° C. Analytical HPLC shows 97.7% purity, (ret time 10.338 min).

Example 221 N-{[6-Chloro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol), 1,1-dimethylethylhexahydro-1H-1,4-diazepine-1-carboxylate (0.012 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in 1,2-dichloroethane (2 mL), and the mixture stirred for thirty min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the protected product. This was treated with 10% TFA in CH₂Cl₂ for 3 h. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the title compound as a white solid. LC-MS m/z 763.2 (M+H)⁺, 1.37 min (ret time); mp 116-119° C. Analytical HPLC shows 95.3% purity, (ret time 10.056 min).

Example 222 N-{[6-Chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol), 1,1-dimethylethyl 1-piperazinecarboxylate (0.0112 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in 1,2-dichloroethane (2 mL), and the mixture stirred for thirty min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Some starting material still remained, so additional 1,1-dimethylethyl 1-piperazinecarboxylate (0.004 g, 0.02 mmol) and sodium triacetoxyborohydride (0.005 g, 0.024 mmol) were added and the mixture again stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the protected product. This was treated with 10% TFA in CH₂Cl₂ for 3 h. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 749.1 (M+H)⁺, 1.47 min (ret time); mp 127-130° C. Analytical HPLC shows 98.8% purity, (ret time 10.129 min).

Example 223 N-({6-Chloro-3′-[(1S,4S)-2,5-diazabicyclo [2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.034 g, 0.05 mmol), (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (0.0119 g, 0.06 mmol), and acetic acid (0.0036 g, 0.06 mmol) were combined in 1,2-dichloroethane (2 mL), and the mixture stirred for 30 min. Sodium triacetoxyborohydride (0.0148 g, 0.07 mmol) was added and the mixture stirred overnight at room temperature. Solvents were concentrated, and the residue taken up in EtOAc, washed with 1N NaOH, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to give the protected product. This was treated with 10% TFA in CH₂Cl₂ for 3 h. The product was purified using a Gilson HPLC with a TFA system. The purified compound was taken up in EtOAc, washed 3× with 2.5 N NaOH, 1× with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to afford the title compound as a white solid. LC-MS m/z 761.1 (M+H)⁺, 1.46 min (ret time); mp 145-149° C. Analytical HPLC shows 99.0% purity, (ret time 10.092 min).

Example 224 N-[(5-Chloro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

1,1-Dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (24.98 mg, 0.060 mmol), N-[(3-bromo-5-chlorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (39.2 mg, 0.06 mmol), potassium carbonate (0.025 g, 0.18 mmol), and Pd(Ph₃P)₄ (3.47 mg, 3.00 μmol) were added to a 0.5-2 mL Biotage microwave vial in 1,4-dioxane (1.5 mL) and water (0.5 mL). The vial was capped and the mixture microwaved at normal power in the SmithCreator microwave at 100° C. for 15 min. LC-MS indicated complete reaction. The crude product was partitioned between EtOAc (30 mL) and water (10 mL). The phases were separated, and the organic washed with brine, dried over anhydrous Na₂SO₄ filtered and concentrated to give the protected product. This was taken up in DCM (1.8 mL) and treated with trifluoroacetic acid (0.2 mL). The mixture was stirred under argon for 5 h. LC-MS indicated complete reaction. Solvents were pumped off and the residue taken up in DMSO and purified on the Gilson with the TFA system. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated and the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give the title compound as a white solid. LC-MS m/z 763.1 (M+H)⁺, 1.67 min (ret time); mp 119-121° C. Analytical HPLC shows 100% purity, (ret time 10.534 min).

Example 225 3-{[{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}(methyl)amino]methyl}-N-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}benzamide

To a solution of 1,1-dimethylethyl 4-{[5′-({[(3-{[{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}(methyl)amino]methyl}phenyl) carbonyl]amino}methyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.055 g, 0.066 mmol) in DCM (0.25 mL) was added TFA (0.25 mL) and stirred at room temperature for 30 min before was diluted with DCM, basified with NaHCO₃ (sat. aq.). Organic layer was separated, dried over Na₂SO₄, filtered, concentrated, purified using a Gilson HPLC (with TFA), final product was basified with an amine cartridge to afford 0.015 g (31%) of the title compound. LC-MS m/z 733 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.24 (t, J=7.40 Hz, 3 H) 1.48 (t, J=7.15 Hz, 3 H) 1.70-1.85 (m, J=10.79 Hz, 2 H) 2.01-2.11 (m, J=12.55, 1.76 Hz, 2 H) 2.56 (s, 3 H) 2.83-2.97 (m, J=7.28 Hz, 2 H) 3.46-3.58 (m, J=5.77 Hz, 8 H) 3.59-3.68 (m, 2 H) 3.92-4.00 (m, J=12.05 Hz, 2 H) 4.01-4.26 (m, J=29.11 Hz, 4 H) 4.26-4.35 (m, 1 H) 4.41-4.50 (m, 4 H) 4.61 (s, 2 H) 7.19 (dd, J=10.67, 8.41 Hz, 1 H) 7.39-7.45 (m, 1 H) 7.50-7.60 (m, 4 H) 7.61-7.70 (m, 2 H) 7.74 (s, 1 H) 7.86 (d, J=8.03 Hz, 1 H) 7.95 (s, 1 H) 8.25 (s, 1 H).

Example 226 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N-methyl-1,3-benzenedicarboxamide

To a solution of 1,1-dimethylethyl 4-{[5′-({[(3-{[{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}(methyl)amino]carbonyl}phenyl) carbonyl]amino}methyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.056 g, 0.066 mmol) in DCM (0.25 mL) was added TFA (0.25 mL) which was then stirred at room temperature for 30 min before being diluted with DCM and basified with NaHCO₃ (sat. aq.). The organic layer was separated, dried over Na₂SO₄, filtered, concentrated, purified using a Gilson HPLC (with TFA), and the final product was basified with an amine cartridge to afford 0.047 g (95%) of the title compound. LC-MS m/z 747 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.39 (t, J=7.53 Hz, 3 H) 1.50 (t, J=7.28 Hz, 3 H) 1.79-1.92 (m, 2 H) 2.08-2.17 (m, J=12.80, 1.76 Hz, 2 H) 2.93 (s, 3 H) 3.15 (q, J=7.45 Hz, 2 H) 3.56 (s, 8 H) 3.62-3.71 (m, 2 H) 3.98-4.06 (m, J=8.66, 2.89 Hz, 2 H) 4.33-4.42 (m, 1 H) 4.45-4.54 (m, 4 H) 4.61 (s, 2 H) 7.17 (dd, J=10.54, 8.53 Hz, 1 H) 7.38-7.44 (m, J=5.52, 3.01 Hz, 1 H) 7.49-7.63 (m, 4 H) 7.64-7.70 (m, 2 H) 7.74 (s, 1 H) 7.97-8.03 (m, 2 H) 8.32 (s, 1 H).

Example 227 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-{[{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}(methyl)amino]methyl}benzamide

To a solution of 1,1-dimethylethyl 4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-phenyl}methyl)(methyl)amino]-methyl}-2′-fluoro-3-biphenylyl)methyl]-1-piperazinecarboxylate (0.386 g, 0.463 mmol) in DCM (3.6 mL) was added TFA (1.8 mL) and this mixture was stirred at room temperature for 100 min before being quenched with NaHCO₃ (sat. aq.). This mixture was extracted with DCM, concentrated, purified using a Gilson HPLC (with TFA). Product was basified with an amine cartridge to afford 0.068 g (20%) of the title compound. LC-MS m/z 733 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.40 (t, J=7.65 Hz, 3 H) 1.49 (t, J=7.15 Hz, 3 H) 1.79-1.92 (m, 2 H) 2.05-2.14 (m, 2 H) 2.74 (s, 3 H) 3.18 (q, J=7.61 Hz, 2 H) 3.43-3.58 (m, 8 H) 3.63-3.72 (m, 2 H) 3.99-4.07 (m, 2 H) 4.30-4.62 (m, 9 H) 4.72 (s, 2 H) 7.34 (dd, J=10.42, 8.66 Hz, 1 H) 7.54-7.65 (m, 4 H) 7.67-7.78 (m, 4 H) 8.02 (d, J=8.03 Hz, 1 H) 8.12 (s, 1 H) 8.29 (s, 1 H).

Example 228 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)methyl]-N-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}benzamide

To a solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.070 g, 0.127 mmol) in DCM (1.5 mL) was added 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.0385 g, 0.127 mmol), NaOH (0.38 mL, 0.5 M aqueous, 0.191 mmol). The mixture was stirred at room temperature for 4 h whereupon NaI (0.019 g, 0.127 mmol) was added. This mixture was stirred for 66 h, at which time added BnN(n-Bu)₃.Br (0.0291 g, 0.127 mmol) was added and that mixture stirred at room temperature for 3 h. Then it was heated in a microwave oven at 60° C. for 30 min, then 1 h. The reaction mixture was concentrated and purified using a Gilson HPLC (w/TFA). The HPLC Fractions containing product were combined and concentrated on a GeneVav EZ-2 evaporator at 45° C. (de-Boc completed during evaporation) and basified with an amine cartridge to afford 0.0133 g (15%) of the title compound. LC-MS m/z 719 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.23 (t, J=7.53 Hz, 3 H) 1.49 (t, J=7.28 Hz, 3 H) 1.84-1.97 (m, J=11.80 Hz, 2 H) 2.02-2.11 (m, J=12.80 Hz, 2 H) 2.82 (q, J=7.53 Hz, 2 H) 3.38-3.47 (m, 4 H) 3.47-3.55 (m, J=4.27 Hz, 4 H) 3.60-3.70 (m, 2 H) 4.03 (dd, J=11.54, 3.01 Hz, 2 H) 4.30-4.40 (m, 3 H) 4.42-4.52 (m, 6 H) 4.62 (s, 2 H) 7.18 (dd, J=10.67, 8.41 Hz, 1 H) 7.37-7.45 (m, J=4.64, 2.38 Hz, 1 H) 7.49-7.67 (m, 5 H) 7.70-7.79 (m, 2 H) 7.91-7.98 (m, 1 H) 8.07 (s, 1 H) 8.30 (s, 1 H).

Example 229 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}oxy)methyl]-N-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}benzamide

To a solution of 5-(chloromethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (73.9 mg, 0.229 mmol) in DMF (0.5 mL) was added 1,1-dimethylethyl 4-({2′-fluoro-5′-[({[3-(hydroxymethyl)phenyl]carbonyl}-amino)methyl]-3-biphenylyl}methyl)-1-piperazinecarboxylate (122 mg, 0.229 mmol). The mixture was stirred at room temperature for 15 h then more 5-(chloro-methyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (73.9 mg, 0.229 mmol) was added. The mixture was stirred for another 24 h. The reaction was quenched with H₂O (1 drop), diluted with MeOH (0.5 mL), purified using a Gilson HPLC (with TFA). To the Fractions containing the product was added TFA (1 drop in each tube ˜12 mL) and solvent was concentrated on a GeneVav EZ-2 evaporator. The compound was purified again with a Gilson HPLC (with TFA) and concentrated on GeneVav EZ-2 evaporator. Residue was basified with an amine cartridge to afford 0.0095 g (6%) of the title compound. LC-MS m/z 720 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.29 (t, J=7.65 Hz, 3 H) 1.50 (t, J=7.28 Hz, 3 H) 1.62-1.75 (m, 2 H) 1.97-2.06 (m, 2 H) 2.96 (q, J=7.53 Hz, 2 H) 3.38-3.47 (m, 4 H) 3.47-3.54 (m, J=4.52 Hz, 4 H) 3.56-3.64 (m, 2 H) 3.85-3.93 (m, 2 H) 4.25-4.33 (m, 1 H) 4.36 (s, 2 H) 4.47 (q, J=7.28 Hz, 2 H) 4.60 (s, 2 H) 4.70 (s, 2 H) 4.83 (s, 2 H) 7.19 (dd, J=10.67, 8.41 Hz, 1H) 7.37-7.43 (m, 1 H) 7.46-7.59 (m, 5 H) 7.65 (d, J=6.78 Hz, 1 H) 7.72 (s, 1 H) 7.81 (d, J=7.78 Hz, 1 H) 7.89 (s, 1 H) 8.27 (s, 1 H).

Example 230 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(2-fluoro-3-formylphenyl)methyl]-1,3-benzenedicarboxamide (60 mg, 0.091 mmol) in DCM (2.0 mL) was added 1-methylpiperazine (0.0202 mL, 0.182 mmol), and NaBH(OAc)₃ (38.6 mg, 0.182 mmol). The mixture was stirred at room temperature for 2 h before being quenched with H₂O (3˜5 drops). The mixture was concentrated and purified using a Gilson HPLC (with TFA). Product fractions were basified with an amine cartridge to afford 0.0509 g (75%) of the title compound. LC-MS m/z 747 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.41 (t, J=7.53 Hz, 3 H) 1.50 (t, J=7.15 Hz, 3 H) 1.83-1.94 (m, 2 H) 2.05-2.15 (m, J=12.55 Hz, 2 H) 2.91 (s, 3 H) 3.04-3.24 (m, J=7.36, 7.36, 7.36 Hz, 6 H) 3.35-3.50 (m, 4 H) 3.62-3.73 (m, 2 H) 4.00-4.09 (m, 4H) 4.31-4.41 (m, 1H) 4.47 (q, J=7.28 Hz, 2 H) 4.67-4.76 (m, J=8.28 Hz, 4 H) 7.24 (t, J=7.65 Hz, 1 H) 7.37-7.48 (m, 3 H) 7.51 (t, J=7.53 Hz, 1 H) 7.55-7.67 (m, 3 H) 8.06 (dd, J=7.91, 1.63 Hz, 2 H) 8.30 (s, 1 H) 8.40 (t, J=1.63 Hz, 1 H).

Example 231 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N,N′-dimethyl-1,3-benzenedicarboxamide

To a solution of 1,6-diethyl-5-[(methylamino)methyl]-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.0416 g, 0.131 mmol) in DMF (1.0 mL) was added 3-{[({4′-[((3S)-4-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl) (methyl)amino]carbonyl}benzoic acid (0.063 g, 0.109 mmol), HBTU (0.0622 g, 0.164 mmol), and Et₃N (0.0306 mL, 0.218 mmol) and this mixture was stirred at room temperature for 64 h before being quenched with H₂O. The mixture was then diluted with MeOH and purified using a Gilson HPLC (with TFA). TFA (1 drop in each tube ˜12 mL) was added to each product-containing tube and solvent removed on GeneVav EZ-2 evaporator. The compound was purified again using a Gilson HPLC (with TFA) and removing solvent using a GeneVav EZ-2 evaporator. Residue was run through an amine cartridge to afford 0.0228 g (27%) of the title compound. LC-MS m/z 775 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.33-1.45 (m, J=6.53 Hz, 6 H) 1.51 (t, J=7.28 Hz, 3 H) 1.75-1.92 (m, 2 H) 2.04-2.19 (m, J=9.29 Hz, 2 H) 2.81-3.05 (m, 5 H) 3.06-3.19 (m, 4 H) 3.23-3.30 (m, 1 H) 3.46 (t, J=13.05 Hz, 1 H) 3.58-3.77 (m, 6 H) 4.02 (dd, J=8.53, 3.01 Hz, 2 H) 4.31-4.46 (m, 3 H) 4.50 (q, J=7.11 Hz, 2 H) 4.57-4.68 (m, 1 H) 4.76-4.84 (m, 1 H) 4.85-4.93 (m, 2 H) 7.16-7.82 (m, 11 H) 8.32 (s, 1 H).

Example 232 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N,N′-dimethyl-1,3-benzenedicarboxamide

To a solution of 1,6-diethyl-5-[(methylamino)methyl]-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.0381 g, 0.12 mmol) in DCM (1.0 mL) was added 3-{[({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-(methyl)amino]carbonyl}benzoic acid (0.0476 g, 0.10 mmol), HBTU (0.0569 g, 0.15 mmol), and Et₃N (0.0281 mL, 0.2 mmol). This mixture was stirred at room temperature for 64 h and then the reaction was quenched with H₂O. The mixture was concentrated, purified using a Gilson HPLC (with TFA), and basified using an amine cartridge to afford 0.0445 g (57%) of the title compound. LC-MS m/z 775 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.39 (t, J=6.78 Hz, 3 H) 1.51 (t, J=7.15 Hz, 3 H) 1.75-1.92 (m, 2 H) 2.04-2.17 (m, 2 H) 2.81-3.05 (m, 8 H) 3.06-3.23 (m, 7 H) 3.36-3.62 (m, 4 H) 3.62-3.72 (m, 2 H) 3.93-4.13 (m, 4 H) 4.31-4.44 (m, 1 H) 4.49 (q, J=7.11 Hz, 2 H) 4.61 (s, 1 H) 4.80 (s, 1 H) 4.92-4.99 (m, J=14.31 Hz, 2 H) 7.15-7.32 (m, 2 H) 7.40-7.69 (m, 9 H) 8.33 (s, 1 H).

Example 233 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-benzenedicarboxamide

To a solution of 4-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino) carbonyl]benzoic acid (0.0452 g, 0.10 mmol) in DCM (1.0 mL) was added 1,1-dimethylethyl (2S)-4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-2-methyl-1-piperazine carboxylate (0.0414 g, 0.10 mmol), HBTU (0.0569 g, 0.15 mmol), and Et₃N (0.0281 mL, 0.20 mmol). This mixture was stirred at room temperature for 1 h, concentrated, and purified using a Gilson HPLC (with TFA). TFA (1 drop in each tube ˜12 mL), was added to Fractions containing product and solvent was concentrated using a GeneVav EZ-2 evaporator. The compound was purified again using a Gilson HPLC (with TFA) and evaporating the solvent using a GeneVav EZ-2 evaporator. Then product was basified with an amine cartridge to afford 0.0234 g (31%) of the title compound. LC-MS m/z 747 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.33 (d, J=6.78 Hz, 3 H) 1.41 (t, J=7.53 Hz, 3 H) 1.50 (t, J=7.15 Hz, 3 H) 1.80-1.97 (m, 2 H) 2.06-2.15 (m, J=13.05 Hz, 2 H) 2.57-2.69 (m, 1 H) 2.75-2.88 (m, J=15.06 Hz, 1 H) 3.11-3.20 (m, 2 H) 3.33-3.39 (m, 3 H) 3.46-3.56 (m, J=13.80 Hz, 2 H) 3.64-3.74 (m, 2 H) 4.00-4.10 (m, J=3.01 Hz, 4 H) 4.31-4.42 (m, 1 H) 4.47 (q, J=7.19 Hz, 2 H) 4.58-4.64 (m, 2 H) 4.70-4.77 (m, 2 H) 7.18 (dd, J=10.54, 8.53 Hz, 1 H) 7.37-7.42 (m, 1 H) 7.43-7.47 (m, 1 H) 7.48-7.54 (m, 2 H) 7.54-7.59 (m, 1 H) 7.62-7.66 (m, 1 H) 7.93-8.01 (m, 4 H) 8.30 (s, 1 H).

Example 234 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,4-benzenedicarboxamide

To a solution of 4-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino) carbonyl]benzoic acid (0.0452 g, 0.10 mmol) in DCM (1.0 mL) was added 1,1-dimethylethyl 4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-1-piperazinecarboxylate (0.040 g, 0.10 mmol), HBTU (0.0569 g, 0.15 mmol), and Et₃N (0.0281 mL, 0.20 mmol). This mixture was stirred at room temperature for 1 h, was concentrated and purified using a Gilson HPLC (with TFA). TFA (1 drop in each tube ˜12 mL) was added to Fractions containing product and solvent was concentrated using a GeneVav EZ-2 evaporator. The compound was purified again using a Gilson HPLC (with TFA) and evaporating the solvent using a GeneVav EZ-2 evaporator. Product was basified with an amine cartridge to afford 0.0284 g (39%) of the title compound. LC-MS m/z 733 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.41 (t, J=7.65 Hz, 3 H) 1.49 (t, J=7.28 Hz, 3 H) 1.81-1.93 (m, 2 H) 2.06-2.14 (m, 2 H) 3.11-3.28 (m, 6 H) 3.39-3.46 (m, J=4.52 Hz, 4 H) 3.64-3.74 (m, 2 H) 4.01-4.10 (m, 2 H) 4.14-4.20 (m, 2 H) 4.32-4.42 (m, J=4.27, 4.27 Hz, 1 H) 4.47 (q, J=7.11 Hz, 2 H) 4.61 (s, 2 H) 4.70-4.76 (m, 2 H) 7.18 (dd, J=10.54, 8.53 Hz, 1 H) 7.37-7.43 (m, 1 H) 7.46-7.56 (m, 3 H) 7.58-7.63 (m, 1 H) 7.67 (s, 1 H) 7.93-8.01 (m, 4 H) 8.30 (s, 1 H).

Example 235 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(2-oxo-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridine-5-yl]methyl}-N′-{[6-fluoro-3′-(hydroxymethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (0.0997 g, 0.15 mmol) in DCM (1.0 mL) was added iodine (0.0381 g, 0.15 mmol), and PS-PPh₃ (0.0818 g, 0.18 mmol). This mixture was stirred at room temperature for 28 h, filtered and concentrated to afford N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(iodomethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide which was carried on to the next step of the reaction directly.

To a solution of 1,1-dimethylethyl 3-oxo-1-piperazinecarboxylate (0.027 g, 0.135 mmol) in DMF (0.25 mL) was added NaH (0.0033 g, 0.135 mmol). This mixture was stirred at room temperature for 5 min whereupon a solution of the above N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(iodomethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide in DMF (0.25 mL) was added. The resulting mixture was stirred for 15 h and purified with a Gilson HPLC (with TFA). The relevant HPLC fractions collected and treated with TFA (0.10 mL), the solvent concentrated on GeneVav EZ-2 evaporator, purified again with Gilson HPLC (with TFA), solvent concentrated on GeneVav EZ-2 evaporator, and the residue basified with an amine cartridge to afford 0.0143 g (13%) of the title compound. LC-MS m/z 747 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.40 (t, J=7.65 Hz, 3 H) 1.49 (t, J=7.15 Hz, 3 H) 1.81-1.93 (m, 2 H) 2.09 (d, J=12.55 Hz, 2 H) 3.16 (q, J=7.53 Hz, 2 H) 3.48-3.55 (m, 2 H) 3.55-3.61 (m, 2 H) 3.62-3.72 (m, 2 H) 3.93 (s, 2 H) 3.99-4.08 (m, 2 H) 4.30-4.40 (m, 1 H) 4.47 (q, J=7.11 Hz, 2 H) 4.60 (s, 2 H) 4.72 (d, J=4.77 Hz, 4 H) 7.15 (dd, J=10.67, 8.41 Hz, 1 H) 7.32-7.41 (m, 2 H) 7.44 (t, J=7.53 Hz, 1 H) 7.47-7.54 (m, 3 H) 7.60 (t, J=7.91 Hz, 1 H) 8.00-8.08 (m, 2 H) 8.29 (s, 1 H) 8.38 (t, J=1.63 Hz, 1 H).

Example 236 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

236(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (130 mg, 0.19 mmol) in DCM (2 mL) was added tert-butyl 1-homopiperazine carboxylate (0.04 mL, 0.21 mmol), AcOH (0.01 mL, 0.23 mmol) followed by NaBH(OAc)₃ (57.2 mg, 0.27 mmol). The reaction mixture was stirred at RT overnight. LC-MS showed minor amount of starting material remaining so 0.5 equiv. of tert-butyl 1-homopiperazine carboxylate and 1 equiv of NaBH(OAc)₃ were added to the mixture. It was stirred for an additional 4 h. LC-MS showed no more starting material. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. It was purified with CombiFlash chromatography, eluting with 0 to 10% MeOH (2M NH₃ in MeOH) in DCM. The product fractions were combined and concentrated under vacuum to give 1,1-dimethylethyl 4-{[3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)amino]methyl}-5′-(methyloxy)-3-biphenylyl]methyl}hexahydro-1H-1,4-diazepine-1-carboxylate (131 mg, 79%). LC-MS m/z 859 (M+H)⁺.

236(b) A mixture of 1,1-dimethylethyl 4-{[3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)-amino]methyl}-5′-(methyloxy)-3-biphenylyl]methyl}hexahydro-1H-1,4-diazepine-1-carboxylate (130 mg, 0.19 mmol) in 25% TFA in DCM (3 mL) was stirred at RT for 2 h. It was purified by preparative hplc (NH₂OH condition), eluting with 10 to 90% CH₃CN in water. The product-containing fractions were dried under GeneVac to give a solid. It was purified using a Gilson HPLC (with 0.1% TFA condition) and eluting with 10 to 90% CH₃CN in water. Product-containing fractions were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (45 mg, 31%). LC-MS m/z 759 (M+H)⁺.

Example 237 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

237(a) A mixture of N-{[3-bromo-5-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (150 mg, 0.23 mmol), {3-[((3S)-4-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]phenyl}boronic acid (100 mg, 0.30 mmol), Na₂CO₃ (74 mg, 0.69 mmol), Pd(PPh₃)₄ (13.3 mg, 0.01 mmol), water (0.75 mL) in dioxane (2.25 mL) was degassed for 5 min. and then heated in a microwave for 30 min at 150° C. The reaction was quenched with water and then extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a 0.45 μM filter and then concentrated to give a crude residue. It was then purified using Combi Flash chromatography, eluting with 0 to 10% MeOH in DCM. The product fractions were combined and concentrated under vacuum to give 1,1-dimethylethyl (2S)-4-{[3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)amino]methyl}-5′-(methyloxy)-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate as a solid (130 mg, 65%): LC-MS m/z 859 (M+H)⁺.

237(b) A mixture of 1,1-dimethylethyl (2S)-4-{[3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}-carbonyl)amino]-methyl}-5′-(methyloxy)-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate (130 mg, 0.19 mmol) in 25% TFA in DCM (3 mL) was stirred at RT for 2 h. It was purified by preparative hplc (NH₂OH condition), eluting with 10 to 90% CH₃CN in water. The fractions were dried under GeneVac to give a solid. It was purified further using a Gilson HPLC (with 0.1% TFA condition) and eluting with 10 to 90% CH₃CN in water. Fractions containing compound were combined and then converted to its free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a white solid (58 mg, 50%). LC-MS m/z 759 (M+H)⁺.

Example 238 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

238(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (130 mg, 0.19 mmol) in DCM (2 mL) was added tert-butyl piperazine (53.8 mg, 0.29 mmol) and AcOH (0.01 mL, 0.23 mmol) followed by NaBH(OAc)₃ (81.7 mg, 0.39 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue: LC-MS m/z 845 (M+H)⁺.

238(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition), eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (58 mg, 40%). LC-MS m/z 745 (M+H)⁺.

Example 239 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (130 mg, 0.19 mmol) in DCM (2 mL) was added 1-methyl piperazine (28.9 mg, 0.29 mmol), AcOH (0.01 mL, 0.23 mmol) followed by NaBH(OAc)₃ (81.7 mg, 0.39 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. The crude residue was purified using a Gilson HPLC (with 0.1% TFA condition), eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give n-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide as a white solid (68 mg, 46%). LC-MS m/z 759 (M+H)⁺.

Example 240 N-{[3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-5-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (130 mg, 0.19 mmol) in DCM (2 mL) was added tert-butyl (1S,4S)-(−)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (42.0 mg, 0.21 mmol), and AcOH (0.01 mL, 0.23 mmol) followed by NaBH(OAc)₃ (57.2 mg, 0.27 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue: LC-MS m/z 857 (M+H)⁺. The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-5-(methyloxy)-3-biphenylyl]methyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a white solid (47 mg, 40%). LC-MS m/z 757 (M+H)⁺.

Example 241 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (130 mg, 0.19 mmol) in DCM (2 mL) was added 2,6-dimethylpiperazine (predominantly cis, 24.2 mg, 0.21 mmol), AcOH (0.01 mL, 0.23 mmol) followed by NaBH(OAc)₃ (57.2 mg, 0.27 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. The crude residue was purified using a Gilson HPLC (with 0.1% TFA condition), eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (75 mg, 50%). LC-MS m/z 773 (M+H)⁺.

Example 242 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-methyl-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

242(a) A mixture of N-[(3-bromo-5-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (150 mg, 0.24 mmol), {3-[((3S)-4-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]phenyl}boronic acid (128 mg, 0.31 mmol), Na₂CO₃ (75 mg, 0.71 mmol), Pd(PPh₃)₄ (13.7 mg, 0.01 mmol), and water (0.75 mL) in dioxane (2.25 mL) was degassed for 5 min. and then heated under microwave for 30 min at 150° C. It was quenched with water and then extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a 0.45 μM filter and then concentrated to give crude residue. It was then purified with Combi Flash chromatography eluting with 0 to 10% MeOH in DCM. The product fractions were combined and concentrated under vacuum to give a crude residue (76 mg, 43%): LC-MS m/z 843 (M+H)⁺.

242(b) The crude product was dissolved in 25% TFA in DCM (3 mL) was stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-methyl-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a white solid (76 mg, 43%). LC-MS m/z 744 (M+H)⁺.

Example 243 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-5-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

243(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added tert-butyl 1-homopiperazine carboxylate (0.04 mL, 0.23 mmol), AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue: LC-MS m/z 844 (M+H)⁺.

243(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-5-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (48 mg, 43%). LC-MS m/z 744 (M+H)⁺.

Example 244 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added 2,6-dimethylpiperazine (predominantly cis) (26 mg, 0.23 mmol), and AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a white solid (50 mg, 43%). LC-MS m/z 757 (M+H)⁺.

Example 245 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

245(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added tert-butyl 1-piperazine carboxylate (42.4 mg, 0.23 mmol), AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue: LC-MS m/z 830 (M+H)⁺.

245(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition), eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (57 mg, 52%). LC-MS m/z 730 (M+H)⁺.

Example 246 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added 1-methyl piperazine (22.8 mg, 0.23 mmol), and AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO3 and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide as a white solid (42 mg, 37%). LC-MS m/z 743 (M+H)⁺.

Example 247 N-({3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-5-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

247(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added tert-butyl (1S,4S)-(−)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (45.1 mg, 0.23 mmol), AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue: LC-MS m/z 842 (M+H)⁺.

247(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-5-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a white solid (49 mg, 44%). LC-MS m/z 742 (M+H)⁺.

Example 248 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(4-methyl-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

248(a) A mixture of N-[(5-bromo-2-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (150 mg, 0.24 mmol), 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (128 mg, 0.31 mmol), Na₂CO₃ (75 mg, 0.71 mmol), Pd(PPh₃)₄ (14 mg, 0.01 mmol), and water (0.75 mL) in dioxane (2.25 mL) was degassed for 5 min. and then heated under microwave for 30 min at 150° C. It was quenched with water and then extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a 0.45 μ syringe filter to get rid of Pd content and then concentrated to give crude residue: LC-MS m/z 844 (M+H)⁺.

248(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at room temperature for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(4-methyl-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a white solid (24 mg, 14%). LC-MS m/z 744 (M+H)⁺.

Example 249 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-4-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added tert-butyl 1-homopiperazine carboxylate (0.04 mL, 0.23 mmol), AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. LC-MS m/z 844 (M+H)⁺.

The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give n-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-4-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (27 mg, 24%). LC-MS m/z 744 (M+H)⁺.

Example 250 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added 2,6-dimethylpiperazine (predominantly cis) (26 mg, 0.23 mmol), AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a white solid (23 mg, 20%). LC-MS m/z 757 (M+H)⁺.

Example 251 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[4-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

251(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added tert-butyl 1-piperazine carboxylate (42.4 mg, 0.23 mmol), AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. LC-MS m/z 829 (M+H)⁺.

251 (b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[4-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (25 mg, 22%). LC-MS m/z 730 (M+H)⁺.

Example 252 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({4-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added 1-methyl piperazine (22.8 mg, 0.23 mmol), and AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({4-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide as a white solid (33 mg, 29%). LC-MS m/z 744 (M+H)⁺.

Example 253 N-({3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-4-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

253(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.15 mmol) in DCM (2 mL) was added tert-butyl (1S,4S)-(−)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (45.1 mg, 0.23 mmol), and AcOH (0.01 mL, 0.18 mmol) followed by NaBH(OAc)₃ (64.3 mg, 0.30 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. LC-MS m/z 842 (M+H)⁺.

253(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 90% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated. However, proton NMR showed minor impurities were present in this batch. It was triturated with ether to give N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-4-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a white solid (18 mg, 16%). LC-MS m/z 742 (M+H)⁺.

Example 254 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide and N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3-fluorophenyl)methyl]-1,3-benzenedicarboxamide

254(a) A mixture of N-[(3-bromo-5-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (200 mg, 0.31 mmol), {3-[((3S)-4-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]phenyl}boronic acid (170 mg, 0.41 mmol), Na₂CO₃ (100 mg, 0.94 mmol), Pd(PPh₃)₄ (18.1 mg, 0.02 mmol), water (0.75 mL) in dioxane (2.25 mL) was degassed for 5 min. and then heated under microwave for 30 min at 150° C. It was quenched with water and then extracted with ethyl acetate twice. The combined organic layers were washed with water and brine. The organic layer was filtered though a 0.45 μM filter and then concentrated to give crude residue. LC-MS m/z 847 (M+H)⁺.

254(b) The crude product was dissolved in 25% TFA in DCM (3 mL) was stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 70% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a solid (53 mg, 23%). LC-MS m/z 747 (M+H)⁺. The biproduct, N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3-fluorophenyl)methyl]-1,3-benzenedicarboxamide (56 mg, 32%), was also isolated as a solid. LC-MS m/z 559 (M+H)⁺.

Example 255 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

255(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (150 mg, 0.23 mmol) in DCM (2 mL) was added tert-butyl 1-piperazine carboxylate (63.2 mg, 0.34 mmol), AcOH (0.02 mL, 0.27 mmol) followed by NaBH(OAc)₃ (95.9 mg, 0.45 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO3 and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. LC-MS m/z 833 (M+H)⁺.

255(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 70% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (33 mg, 30%). LC-MS m/z 733 (M+H)⁺.

Example 256 N-({3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-5-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

256(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (150 mg, 0.23 mmol) in DCM (2 mL) was added tert-butyl (1S,4S)-(−)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (67.3 mg, 0.34 mmol), AcOH (0.02 mL, 0.27 mmol) followed by NaBH(OAc)₃ (95.9 mg, 0.45 mmol). The reaction mixture was stirred at RT overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. LC-MS m/z 845 (M+H)⁺.

256(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 70% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-5-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a white solid (28 mg, 25%). LC-MS m/z 745 (M+H)⁺.

Example 257 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-fluoro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

257(a) To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (90 mg, 0.14 mmol) in DCM (2 mL) was added tert-butyl 1-homopiperazine carboxylate (0.04 mL, 0.20 mmol), AcOH (0.01 mL, 0.16 mmol) followed by NaBH(OAc)₃ (57.6 mg, 0.27 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give a crude residue. LC-MS m/z 847 (M+H)⁺.

257(b) The crude residue was dissolved in 25% TFA in DCM (2 mL) and stirred at RT for 2 h. It was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 70% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-fluoro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide as a white solid (27 mg, 24%). LC-MS m/z 747 (M+H)⁺.

Example 258 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (90 mg, 0.14 mmol) in DCM (2 mL) was added 2,6-dimethylpiperazine (predominantly cis) (23.3 mg, 0.20 mmol), AcOH (0.01 mL, 0.16 mmol) followed by NaBH(OAc)₃ (57.6 mg, 0.27 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give the crude residue. The crude residue was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 70% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give n-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a solid (25 mg, 22%). LC-MS m/z 761 (M+H)⁺.

Example 259 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (90 mg, 0.14 mmol) in DCM (2 mL) was added 1-methyl piperazine (20.4 mg, 0.20 mmol), AcOH (0.01 mL, 0.16 mmol) followed by NaBH(OAc)₃ (57.6 mg, 0.27 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO₃ and extracted with DCM twice. The combined organic layers were washed with brine and then concentrated under vacuum to give the crude residue. The crude residue was purified using a Gilson HPLC (with 0.1% TFA condition) eluting with 10 to 70% CH₃CN in water at a flowrate of 20 mL/min. Fractions containing product were combined and then converted to the free base with saturated NaHCO₃. The organic layer was dried over sodium sulfate, filtered and then concentrated to give n-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide as a solid (26 mg, 23%). LC-MS m/z 747 (M+H)⁺.

Example 260 N-{[6-Cyano-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Cyano-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (129 mg, 0.000193 mol) in DCM (3 mL) was treated with 1,1-dimethylethyl 1-piperazinecarboxylate (53.8 mg, 0.00028 mol) followed by sodium triacetoxyborohydride (81.7 mg, 0.000385 mol) and AcOH (13.9 mg, 13 μL). Solution was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl, dried (Na₂SO₄) and concentrated giving a glassy foam (173.4 mg). The glassy foam was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. The solution was stripped to dryness. Purification was carried out on a Gilson Instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined, diluted with EtOAc, washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a chalky, white solid [78.2 mg (55%)]. LC-MS m/z 740.5 (M+H)⁺.

Example 261 N-[(6-Cyano-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

261(a) N-[(3-Bromo-4-cyanophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (96.6 mg, 0.00015 mol) in dioxane (4 mL) and 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (81.2 mg, 0.000195 mol) were treated with Na₂CO₃ (48 mg, 0.00045 mol) and 0.75 mL H₂O. The mixture was degassed for 15 min with N₂, then treated with tetrakistriphenylphosphine palladium (0)° (17.3 mg, 0.000015 mol). The reaction was microwaved at 150° C. for 30 min. The reaction was complete by LC-MS. The reaction was quenched with H₂O, then diluted and extracted with EtOAc (2×). Combined organic extract were washed with H₂O, sat. aq. NaCl and dried (Na₂SO₄) and concentrated to give 206.6 mg crude t-BOC product.

261 (b) The crude product was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. The solution was stripped off. Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase back-extracted with EtOAc. Combined organic layers were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated. The residue was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a pink, white solid [66.1 mg (58%)]. LC-MS m/z 754.3 (M+H)⁺.

Example 262 N-({6-Cyano-3′-[(1S,4S)-2,5-diazabicyclo [2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Cyano-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (108 mg, 0.000161 mol) in DCM (4 mL) and 1,1-dimethylethyl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (46 mg, 0.00023 mol) were mixed with sodium triacetoxyborohydride (68.3 mg, 0.00032 mol) and AcOH (11.6 mg, 11 μL). The solution was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). The combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a glassy foam (227.5 mg). The glassy foam was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient on a C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase back-extracted with EtOAc. Combined organic layers were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a chalky, white solid [71.5 mg (59%)]. LC-MS m/z 752.3 (M+H)⁺.

Example 263 N-{[6-Cyano-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Cyano-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (108 mg, 0.000161 mol) in DCM (4 mL) and 1,1-dimethylethyl hexahydro-1H-1,4-diazepine-1-carboxylate (47 mg, 0.00023 mol) were mixed with sodium triacetoxyborohydride (68.3 mg, 0.00032 mol) and AcOH (11.6 mg, 11 μL). This solution was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined extracts were washed with sat. aq. NaCl, dried (Na₂SO₄) and concentrated to a glassy foam (238.7 mg). The glassy foam was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic layers were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a chalky solid [73 mg (60%)]. LC-MS m/z 754.3 (M+H)⁺.

Example 264 N-[(6-Cyano-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Cyano-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (108 mg, 0.000161 mol) in DCM (4 mL) and (2R,6S)-2,6-dimethylpiperazine (26.7 mg, 0.00023 mol) were mixed with sodium triacetoxyborohydride (68.3 mg, 0.00032 mol) and AcOH (11.6 mg, 11 μL). This solution was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). The combined organic extracts were washed with sat. aq. NaCl, dried (Na₂SO₄) and concentrated to a glassy foam (122.9 mg). Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic layers were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a white solid [62.3 mg (51%)]. LC-MS m/z 769.7 (M+H)⁺.

Example 265 N-({6-Cyano-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Cyano-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (108 mg, 0.000161 mol) in DCM (4 mL) and 1-methylpiperazine (23.4 mg, 0.00023 mol) were mixed with sodium triacetoxyborohydride (68.3 mg, 0.00032 mol) and AcOH (11.6 mg, 11 μL). The solution was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). The combined organic extracts were washed with sat. aq. NaCl, dried (Na₂SO₄) and concentrated to a glassy foam (98.9 mg). Purification was carried out on Gilson instrument with an acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase back-extracted with EtOAc. The combined organic layers were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a white solid [43.9 mg (36%)]. LC-MS m/z 769.7 (M+H)⁺.

Example 266 N-({6-Cyano-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(6-Cyano-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (182 mg, 0.000272 mol) in DCM (4 mL) and 1-methylhexahydro-1H-1,4-diazepine (43.4 mg, 0.00038 mol) were mixed with sodium triacetoxyborohydride (115 mg, 0.00054 mol) and AcOH (19.5 mg, 19 μL). The solution was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a glassy foam (150 mg). Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic layers were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a solid. Further purification of the material was needed and this was carried out on a Chomatotron utilizing silica gel [DCM-MeOH-conc. aq. NH₃, (90:7:3)] to afford 42 mg of the title compound as a glassy solid. LC-MS m/z 768.7 (M+H)⁺.

Example 267 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[4-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (54 mg, 0.00008 mol) in DCM with 1,1-dimethylethyl 1-piperazinecarboxylate (23.3 mg, 0.00012 mol) and sodium triacetoxyborohydride (34 mg., 0.00016 mol) and AcOH (6 μL) were agitated overnight on a shaker. LC-MS was used to monitor the reaction. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. The residue was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on a Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a white solid [37.6 mg (63%)]. LC-MS m/z 745.4 (M+H)⁺.

Example 268 N-{[3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-4-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (54 mg, 0.00008 mol) in DCM, 1,1-dimethylethyl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (23 mg, 0.00012 mol) and sodium triacetoxyborohydride (34 mg., 0.00016 mol) and AcOH (6 uL) were agitated overnight on a shaker. LC-MS was used to monitor the reaction. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. The residue was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a peach colored solid [40.9 mg (68%)]. LC-MS m/z 757.5 (M+H)⁺.

Example 269 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (54 mg, 0.00008 mol) in DCM, 1,1-dimethylethyl hexahydro-1H-1,4-diazepine-1-carboxylate (24 mg, 0.00012 mol) and sodium triacetoxyborohydride (34 mg., 0.00016 mol) and AcOH (6 uL) were agitated overnight on a shaker. LC-MS was used to monitor the reaction. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. The residue was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a chalky white solid [36.6 mg (60%)]. LC-MS m/z 760 (M+H)⁺.

Example 270 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (54 mg, 0.00008 mol) in DCM, (2R,6S)-2,6-dimethylpiperazine (13.7 mg, 0.00012 mol) and sodium triacetoxyborohydride (34 mg., 0.00016 mol) and AcOH (6 uL) were agitated overnight on a shaker. LC-MS was used to monitor the reaction. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. Purification was carried out on Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a chalky white solid [41.8 mg (67.6%)]. LC-MS m/z 773 (M+H)⁺.

Example 271 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({4-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (54 mg, 0.00008 mol) in DCM, 1-methylpiperazine (12 mg, 13.3 uL, 0.00012 mol) and sodium triacetoxyborohydride (34 mg., 0.00016 mol) and AcOH (6 μL) were agitated overnight on a shaker. LC-MS was used to monitor the reaction. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. Purification was carried out on Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a chalky white solid [36.2 mg (60%)]. LC-MS m/z 759.5 (M+H)⁺.

Example 272 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (54 mg, 0.00008 mol) in DCM, 1-methylhexahydro-1H-1,4-diazepine (13.7 mg, 15.2 μL, 0.00012 mol) and sodium triacetoxyborohydride (34 mg., 0.00016 mol) and AcOH (6 μL) were agitated overnight on a shaker. LC-MS was used to monitor the reaction. The reaction was quenched with sat. aq. NaHCO₃, then extracted with DCM (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. Purification was carried out on Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a chalky white solid. Further purification was carried out on an Agilent HPLC with RP-CH₃CN:H₂O-conc. aq. NH₃ gradient [4.5 mg (7.25%)]. LC-MS m/z 773.5 (M+H)⁺.

Example 273 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(4-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[5-Bromo-2-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (97 mg, 0.00015 mol) in dioxane (4 mL) and 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (81.2 mg, 0.000195 mol) were treated with Na₂CO₃ (48 mg, 0.00045 mol) and 0.75 mL H₂O. The mixture was degassed for 15 min with N₂, and then treated with tetrakistriphenylphosphine palladium (0) (17.3 mg, 0.000015 mol). The reaction was microwaved at 150° C. for 30 min. The reaction was quenched with H₂O, then diluted and extracted with EtOAc (2×). Combined organic extracts were washed with H₂O, sat. aq. NaCl and dried (Na₂SO₄) and concentrated to give 206.6 mg crude t-BOC product. The crude product was taken up in DCM (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as a beige colored solid (16.4 mg, 14%). LC-MS m/z 759.4 (M+H)⁺.

Example 274 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[2-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-2-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.000152 mol) in 1,2-dichloroethane (4 mL), 1,1-dimethylethyl 1-piperazinecarboxylate (44.6 mg, 0.00024 mol) and sodium triacetoxyborohydride (68 mg, 0.00032 mol) and AcOH (12 μL) was placed on an oscillating shaker overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃, then extracted with 1,2-dichloroethane (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. The crude product was taken up in 1,2-dichloroethane (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on a Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of 1,2-dichloroethane and transferred to a vial and pumped down to afford the title compound as a beige colored solid [21 mg (19%)]. LC-MS m/z 729.6 (M+H)⁺.

Example 275 N-({3′-[(1S,4S)-2,5-Diazabicyclo [2.2.1]hept-2-ylmethyl]-2-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

A mixture of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-2-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.000152 mol) in 1,2-dichloroethane (4 mL), 1,1-dimethylethyl 1-piperazinecarboxylate (44.6 mg, 0.00024 mol) and sodium triacetoxyborohydride (68 mg., 0.00032 mol) and AcOH (12 μL) was placed on an oscillating shaker overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃, then extracted with 1,2-dichloroethane (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. The crude product was taken up in 1,2-dichloroethane (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on a Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of 1,2-dichloroethane and transferred to a vial and pumped down to afford the title compound as a beige colored solid [21.4 mg (19%)]. LC-MS m/z 741.7 (M+H)⁺.

Example 276 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-2-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

A mixture of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-2-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.000152 mol) in 1,2-dichloroethane (4 mL) along with 1,1-dimethylethyl hexahydro-1H-1,4-diazepine-1-carboxylate (48 μL, 0.00024 mol) and sodium triacetoxyborohydride (68 mg., 0.00032 mol) and AcOH (12 μL) was placed on an oscillating shaker overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃, then extracted with 1,2-dichloroethane (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. The crude product was taken up in 1,2-dichloroethane (10 mL) containing 25% TFA and allowed to stand at room temperature for 3 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on a Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of 1,2-dichloroethane and transferred to a vial and pumped down to afford the title compound as a pale brown colored solid [40.6 mg (36%)]. LC-MS m/z 743.9 (M+H)⁺.

Example 277 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

A mixture of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-2-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.000152 mol) in 1,2-dichloroethane (4 mL) with (2R,6S)-2,6-dimethylpiperazine (27.4 mg, 0.00024 mol) and sodium triacetoxyborohydride (68 mg., 0.00032 mol) and AcOH (12 μL) was placed on an oscillating shaker overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃, then extracted with 1,2-dichloroethane (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. Purification was carried out on a Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP Column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of 1,2-dichloroethane and transferred to a vial and pumped down to afford the title compound as a pale brown colored solid [45.3 mg (39%)]. LC-MS m/z 757.5 (M+H)⁺.

Example 278 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

A mixture of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-2-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.000152 mol) in 1,2-dichloroethane (4 mL) with 1-methylpiperazine (26.6 mg., 0.00024 mol) and sodium triacetoxyborohydride (68 mg, 0.00032 mol) and AcOH (12 μL) was placed on an oscillating shaker overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃, then extracted with 1,2-dichloroethane (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. Purification was carried out on Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of 1,2-dichloroethane and transferred to a vial and pumped down to afford the title compound as a pale brown colored solid [55.1 mg (49%). LC-MS m/z 743.9 (M+H)⁺.

Example 279 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-methyl-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

A mixture of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-2-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.000152 mol) in 1,2-dichloroethane (4 mL) with 1-methylhexahydro-1H-1,4-diazepine (30.4 μL., 0.00024 mol) and sodium triacetoxyborohydride (68 mg., 0.00032 mol) and AcOH (12 μL) was placed on an oscillating shaker overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃, then extracted with 1,2-dichloroethane (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. Purification was carried out on Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of 1,2-dichloroethane and transferred to a vial and pumped down to afford the title compound as a pale brown colored solid [48.3 mg (42%)]. LC-MS m/z 757.5 (M+H)⁺.

Example 280 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(2-methyl-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

A mixture of N-[(3-bromo-2-methylphenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (93 mg, 0.00015 mol) in dioxane (4 mL) with 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (81.2 mg, 0.000195 mol) was treated with Na₂CO₃ (48 mg, 0.00045 mol) and 0.75 mL H₂O. The mixture was degassed for 15 min with N₂, then treated with (PPh₃)₄Pd (17.3 mg, 0.000015 mol). The mixture was microwaved at 150° C. for 30 min. The reaction was quenched with H₂O, then diluted and extracted with EtOAc (2×) combined organic extracts were washed with H₂O, sat. aq. NaCl and dried (Na₂SO₄) and concentrated to give crude t-Boc product. The crude product was taken up in DCM (10 mL) containing 33% TFA and allowed to stand at room temperature for 1.5 h. LC-MS showed the reaction to be complete. Solvent was stripped off. Purification was carried out on a Gilson instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of DCM and transferred to a vial and pumped down to afford the title compound as an ivory colored glassy solid [65.9 mg (58%)]. LC-MS m/z 743.7 (M+H)⁺.

Example 267 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3R)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

A mixture of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (100 mg, 0.000152 mol) in 1,2-dichloroethane (4 mL) with 1,1-dimethylethyl (2R)-2-methyl-1-piperazinecarboxylate (48 mg, 0.00024 mol) and sodium triacetoxyborohydride (68 mg, 0.00032 mol) and AcOH (12 μL) was placed on an oscillating shaker overnight at room temperature. The reaction was quenched with sat. aq. NaHCO₃, then extracted with 1,2-dichloroethane (2×). Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. The crude product was taken up in 1,2-dichloroethane (10 mL) containing 25% TFA and allowed to stand at room temperature for 1.5 h. LC-MS showed the reaction to be complete and the solution stripped to dryness. Purification was carried out on Gilson HPLC instrument with acetonitrile-water (0.1% TFA) gradient; C18-RP Column. Fractions containing the clean desired product were combined; diluted with EtOAc; washed with sat. aq. NaHCO₃; and the aq. phase was back-extracted with EtOAc. Combined organic extracts were washed with sat. aq. NaCl and dried (Na₂SO₄) and concentrated to a residue. It was taken up in a small amount of 1,2-dichloroethane and transferred to a vial and pumped down to afford the title compound as an ivory colored solid [62.1 mg (53%)]. LC-MS m/z 747.3 (M+H)⁺.

Example 282 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

A mixture of 1,1-dimethylethyl (2S)-4-[(3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}-carbonyl)amino]methyl}-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate (33 mg), TFA (0.2 mL) and DCM (1.0 mL) was stirred at room temperature overnight. Then TEA (0.4 mL) was added at −78° C. and the solution was then concentrated. Dilution with MeOH (0.5 mL) followed by separation via a Gilson HPLC (basic conditions) afforded the desired product (13 mg). LC-MS m/z 729 (M+H)⁺.

Example 283 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(2-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

283(a) A mixture of N-[(3-bromo-2-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (66 mg crude ˜0.1 mmol), {3-[((3S)-4-{[(1,1-dimethylethyl)-oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]phenyl}boronic acid (43 mg, 0.13 mmol), and Na₂CO₃ (32 mg, 0.3 mmol) in dioxane/water (3:1, 4 mL) was bubbled with N₂ for about 5 min and then Pd(PPh₃)₄ (12 mg, 0.01 mmol) was added. The tube was sealed and irradiated with a microwave reactor for 30 min at 120° C. It was mixed with NaHCO₃ (25 mL) and washed with EtOAc (25 mL). Purification via a Gilson HPLC (acidic conditions) then provided a crude mixture of 1,1-dimethylethyl (2S)-4-[(3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-phenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate.

283(b) The partially purified 1,1-dimethylethyl (2S)-4-[(3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-phenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate was mixed with THF (1 mL) and TFA (0.2 mL). The mixture was stirred at room temperature for 1 h. Purification was done with a Gilson HPLC using TFA conditions. The desired fraction was collected, concentrated and re-dissolved in MeOH. Filtration though an amine cartridge and solvent concentration then afforded the above named product (10 mg). LC-MS m/z 747 (M+H)⁺.

Example 284 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(3,5-dimethyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (50.0 mg. 0.076 mmol) was diluted in DMSO (1.0 mL) and dispensed into a 1 dram vial containing 2,6-dimethylpiperazine (0.76 mmol), ZnCl₂ (0.032 mmol, 4.2 mg), NaB(OAc)₃H (0.76 mmol, 180 mg), with fitted magnetic stir bar, for 12 h. Purification was completed via a Gilson HPLC and desired product collected. The product was dissolved in dichloromethane (1 mL) and washed though 2 g of SPE-Amine in a column (EtOAc:MeOH=4:1, 5 mL) to afford 29.9 mg of the title compound (51.6%). LC-MS m/z 762 (M+H)⁺, 1.38 min (ret time).

Example 285 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(2-hydroxyethyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-(2-hydroxyethyl)piperazine (0.76 mmol) in place of 2,6 dimethylpiperazine to afford 30.7 mg (52.0%) of the final product. LC-MS m/z 778 (M+H)⁺, 1.3 min (ret time).

Example 286 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(2,3,5,6-tetramethyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 2,3,5,6-tetramethylpiperazine (0.76 mmol) for 2,6-dimethylpiperazine to afford 13.5 mg (22.4%) of the final product. LC-MS m/z 790 (M+H)⁺, 1.4 min (ret time).

Example 287 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({4-[2-(methyloxy)ethyl]-1-piperazinyl}methyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-(2-methoxyethyl)piperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 28.9 mg (48.0%) of the final product. LC-MS m/z 792 (M+H)⁺, 1.32 min (ret time).

Example 288 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({4-[2-(dimethylamino)ethyl]-1-piperazinyl}methyl)-6-fluoro-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-[2-(dimethylamino)ethyl]piperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 30.5 mg (49.9%) of the final product. LC-MS m/z 805 (M+H)⁺, 1.21 min (ret time).

Example 289 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-phenyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-phenylpiperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 28.6 mg (46.5%) of the final product. LC-MS m/z 810 (M+H)⁺, 1.59 min (ret time).

Example 290 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(4-pyridinyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-(4-pyridyl)piperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 27.1 mg (44.0%) of the final product. LC-MS m/z 811 (M+H)⁺, 1.24 min (ret time).

Example 291 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({methyl[2-(methylamino)ethyl]amino}methyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using N,N′-dimethylethylenediamine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 2.34 mg (4.1%) of the final product. LC-MS m/z 736 (M+H)⁺, 1.18 min (ret time).

Example 292 N-[(3′-{[(Cyclopropylmethyl)amino]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using (aminomethyl)cyclopropane (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 23.7 mg (43.4%) of the final product. LC-MS m/z 718 (M+H)⁺, 1.45 min (ret time).

Example 293 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[methyl(1-methyl-4-piperidinyl)amino]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-methyl-4-(methylamino)piperidine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 28.2 mg (47.9%) of the final product. LC-MS m/z 776 (M+H)⁺, 1.2 min (ret time).

Example 294 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(2,2,6,6-tetramethyl-4-piperidinyl)amino]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 4-amino-2,2,6,6-tetramethylpiperidine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 27.8 mg (45.5%) of the final product. LC-MS m/z 804 (M+H)⁺, 1.24 min (ret time).

Example 295 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(4-morpholinyl)-1-piperidinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 4-morpholinopiperidine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 33.2 mg (53.5%) of the final product. LC-MS m/z 818 (M+H)⁺, 1.21 min (ret time).

Example 296 N-({3′-[(1-Azabicyclo [2.2.2]oct-3-ylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-aza-bicyclo(2.2.2)oct-3-ylamine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 25.9 mg (44.1%) of the title compound. LC-MS m/z 774 (M+H)⁺, 1.22 min (ret time).

Example 297 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(1-ethyl-3-piperidinyl)amino]methyl}-6-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-ethyl-piperidin-3-ylamine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 31.5 mg (53.5%) of the final product. LC-MS m/z 776 (M+H)⁺, 1.2 min (ret time).

Example 298 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({[(3R)-2-oxohexahydro-1H-azepin-3-yl]amino}methyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using (s)-alpha-amino-omega-caprolactam (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 28.1 mg (47.7%) of the final product. LC-MS m/z 776 (M+H)⁺, 1.36 min (ret time).

Example 299 N-[(3′-{[(2S)-2-(Aminocarbonyl)-1-pyrrolidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using L-prolinamide (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 27.4 mg (47.3%) of the final product. LC-MS m/z 762 (M+H)⁺, 1.33 min (ret time).

Example 300 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({2-[(dimethylamino)methyl]-1-piperidinyl}methyl)-6-fluoro-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using N-(2-piperidylmethyl)-dimethylamine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 27.7 mg (47.9%) of the final product. LC-MS m/z 762 (M+H)⁺, 1.25 min (ret time).

Example 301 N-[(3′-{[3-(Aminocarbonyl)-1-piperidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using nipecotamide (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 26.6 mg (45.1%) of the final product. LC-MS m/z 776 (M+H)⁺, 1.34 min (ret time).

Example 302 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(2-pyrimidinyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 2-piperazin-1-yl-pyrimidine dihydrochloride (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 7.9 mg (12.8%) of the final product. LC-MS m/z 812 (M+H)⁺, 1.44 min (ret time).

Example 303 N-({3′-[(4-Acetyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-acetylpiperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 26.4 mg (44.9%) of the final product. LC-MS m/z 776 (M+H)⁺, 1.35 min (ret time).

Example 304 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(1R,4S)-5-(phenylmethyl)-2,5-diazabicyclo [2.2.1]hept-2-yl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using (1S,4S)-2-benzyl-2,5-diazabicyclo[2.2.1]heptane dihydrobromide (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 21.9 mg (34.4%) of the final product. LC-MS m/z 836 (M+H)⁺, 1.42 min (ret time).

Example 305 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({4-[(3-methylphenyl)methyl]-1-piperazinyl}methyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-(3-methyl-benzyl)-piperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 28.7 mg (45.0%) of the final product. LC-MS m/z 838 (M+H)⁺, 1.57 min (ret time).

Example 306 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(4-fluorophenyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-(4-fluorophenyl)piperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 29.3 mg (46.7%) of the final product. LC-MS m/z 828 (M+H)⁺, 1.62 min (ret time).

Example 307 N-({3′-[(Cyclopentylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using cyclopentylamine in place of 2,6-dimethylpiperazine (0.76 mmol) to afford 9.19 mg (16.5%) of the final product. LC-MS m/z 733 (M+H)⁺, 1.51 min (ret time).

Example 308 N-({3′-[(Cyclohexylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using cyclohexylamine in place of 2,6-dimethylpiperazine (0.76 mmol) to afford 23.6 mg (41.6%) of the final product. LC-MS m/z 747 (M+H)⁺, 1.46 min (ret time).

Example 309 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(2-pyridinylamino)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 2-aminopyridine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 23.8 mg (42.3%) of the final product. LC-MS m/z 741 (M+H)⁺, 1.47 min (ret time).

Example 310 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(3-pyridinylamino)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 3-aminopyridine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 20.7 mg (36.7%) of the final product. LC-MS m/z 741 (M+H)⁺, 1.45 min (ret time).

Example 311 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-pyridinylamino)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 4-aminopyridine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 1.69 mg (3.0%) of the final product. LC-MS m/z 741 (M+H)⁺, 1.45 min (ret time).

Example 312 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-pyrimidinylamino) methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 4-aminopyrimidine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 0.34 mg (0.6%) of the final product. LC-MS m/z 742 (M+H)⁺, 1.47 min (ret time).

Example 313 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(2-pyrimidinylamino) methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 2-aminopyrimidine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 5.48 mg (9.7%) of the final product. LC-MS m/z 742 (M+H)⁺, 1.66 min (ret time).

Example 314 N-({3′-[(Diethylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using diethylamine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 20.8 mg (38.0%) of the final product. LC-MS m/z 720 (M+H)⁺, 1.36 min (ret time).

Example 315 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({[2-(dimethylamino)ethyl]amino}-methyl)-6-fluoro-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using N,N-dimethylethylenediamine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 22.5 mg (40.3%) of the final product. LC-MS m/z 736 (M+H)⁺, 1.26 min (ret time).

Example 316 N-[(3′-{[3-(Aminocarbonyl)-2,2,5,5-tetramethyl-1-pyrrolidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 2,2,5,5-tetramethyl-3-pyrrolidinecarboxamide (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 13.6 mg (21.9%) of the final product. LC-MS m/z 818 (M+H)⁺, 1.45 min (ret time).

Example 317 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 284 using 1-methylpiperazine (0.76 mmol) in place of 2,6-dimethylpiperazine to afford 15.9 mg (28.0%) of the final product. LC-MS m/z 747 (M+H)⁺, 1.34 min (ret time).

Example 318 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-piperidinylamino)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (50.0 mg, 0.076 mmol) was diluted in DMSO (1.0 mL) and dispensed into a 1 dram vial, with fitted magnetic stir bar, containing 4-amino-1-Boc-piperidine (0.76 mmol), ZnCl₂ (0.032 mmol, 4.2 mg), NaB(OAc)₃H (0.76 mmol, 180 mg) and the reaction stirred for 12 h. Purification was performed via a Gilson and the desired product was collected. The desired product was placed in a 1 dram vial with dioxane:MeOH (1.5 mL:0.5 mL) and 2 drops of HCl (concentrated) was added. This was heated at 60° C. for 1 h. The solvent was removed and added to dichloromethane (1 mL) and washed though 2 g SPE-Amine column (EtOAc:MeOH=4:1, 5 mL) to afford 28.6 mg of the title compound (50.3%). LC-MS m/z 748 (M+H)⁺, 1.16 min (ret time).

Example 319 N-({3′-[(4-Amino-1-piperidinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 318 using 4-(N-Boc-amino)piperidine (0.76 mmol) in place of 4-amino-1-Boc-piperidine to afford 23.1 mg (40.6%) of the final product. LC-MS m/z 748 (M+H)⁺, 1.27 min (ret time).

Example 320 N-[(3′-{[(3S)-3-Amino-1-pyrrolidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 318 using (S)-3-(Boc-amino)pyrrolidine (0.76 mmol) in place of 4-amino-1-Boc-piperidine to afford 24.7 mg (44.3%) of the final product. LC-MS m/z 734 (M+H)⁺, 1.19 min (ret time).

Example 321 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(4-piperidinylmethyl)amino]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 318 using 1-Boc-4-(aminomethyl)piperidine (0.76 mmol) in place of 4-amino-1-Boc-piperidine to afford 28.9 mg (49.9%) of the final product. LC-MS m/z 762 (M+H)⁺, 1.17 min (ret time).

Example 322 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(hexahydro-1H-azepin-3-ylamino)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 318 using 1,1-dimethylethyl 3-aminohexahydro-1H-azepine-1-carboxylate (0.76 mmol) in place of 4-amino-1-Boc-piperidine to afford 17.4 mg (30.0%) of the final product. LC-MS m/z 762 (M+H)⁺, 1.38 min (ret time).

Example 323 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(hexahydro-1H-azepin-4-ylamino)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 318 using 1,1-dimethylethyl-4-aminohexahydro-1H-azepine-1-carboxylate (0.76 mmol) in place of 4-amino-1-Boc-piperidine to afford 33.2 mg (57.4%) of the final product. LC-MS m/z 762 (M+H)⁺, 1.17 min (ret time) LC-MS: m/z, 762 (M+H)⁺, r.t. 1.17 min.

Example 324 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-2,3-naphthalenedicarboxamide

To 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (0.1 mmol) and 2,3-naphthalenedicarboxylic acid (0.1 mmol) was dissolved in DCM (3 mL) was added HOBt (1.0 eq, 14.0 mg) and EDC (1.0 eq, 19.0 mg). The resultant solution was stirred overnight. The solution was purified by preparative hplc (Gilson) to yield 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-2-naphthalenecarboxylic acid. This compound was dissolved in 3 mL of DCM with 1,1-dimethylethyl (2S)-4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate (41.3 mg, 0.1 mmol) to which was added HOBt (1.0 eq, 14.0 mg) and EDC (1.0 eq, 19.0 mg). The resultant solution was stirred overnight and purified by preparative hplc (Gilson) to give 1,1-dimethylethyl (2S)-4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-2-naphthalenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate. This compound was then dissolved in 2 mL of dioxane:MeOH (3:1). Three drops of HCl (concentrated) were added to the resultant solution and it was heated at 60° C. for 1 h. The solution was applied to an amine column which was rinsed with 10 mL of dioxane:MeOH (3:1) to afford 2.7 mg (3.4%) of the above named product. LC-MS m/z 798 (M+H)⁺, 1.42 min (ret time).

Example 325 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 5-methyl-1,3-benzenedicarboxylic acid (0.1 mmol) in place of 2,3-naphthalenedicarboxylic acid to afford 13.0 mg (17.1%) of the final product. LC-MS m/z 762 (M+H)⁺, 1.36 min (ret time).

Example 326 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-hydroxy-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 5-hydroxy-1,3-benzenedicarboxylic acid in place of 2,3-naphthalenedicarboxylic acid (0.1 mmol) to afford 4.3 mg (5.6%) of the final product. LC-MS m/z 764 (M+H)⁺, 1.30 min (ret time).

Example 327 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-(1,1-dimethylethyl)-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 5-(1,1-dimethylethyl)-1,3-benzenedicarboxylic acid (0.1 mmol) in place of 2,3-naphthalenedicarboxylic acid to afford 10.0 mg (12.5%) of the final product. LC-MS m/z 804 (M+H)⁺, 1.53 min (ret time).

Example 328 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-2,4,5,6-tetrafluoro-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 2,4,5,6-tetrafluoro-1,3-benzenedicarboxylic acid (0.1 mmol) in place of 2,3-naphthalenedicarboxylic acid to afford 22.0 mg (26.5%) of the final product. LC-MS m/z 820 (M+H)⁺, 1.34 min (ret time).

Example 329 5-Bromo-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 5-bromo-1,3-benzenedicarboxylic acid (0.1 mmol) in place of 2,3-naphthalenedicarboxylic acid to afford 11.0 mg (13.3%) of the final product. LC-MS m/z 827 (M+H)⁺, 1.46 min (ret time).

Example 330 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-naphthalenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 1,4-naphthalenedicarboxylic acid (0.1 mmol) in place of 2,3-naphthalenedicarboxylic acid to afford 21.0 mg (26.3%) of the final product. LC-MS m/z 798 (M+H)⁺, 1.34 min (ret time).

Example 331 2,5-Dichloro-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 2,5-dichloro-1,4-benzenedicarboxylic acid (0.1 mmol) in place of 2,3-naphthalenedicarboxylic acid to afford 31.0 mg (38%) of the final product. LC-MS m/z 817 (M+H)⁺, 1.31 min (ret time).

Example 332 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4b]pyridin-5-yl]methyl}-2,3,5,6-tetrafluoro-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-benzenedicarboxamide

The title compound was prepared according to the general procedure of Example 324 using 2,3,5,6-tetrafluoro-1,4-benzenedicarboxylic acid (0.1 mmol) in place of 2,3-naphthalenedicarboxylic acid to afford 24.6 mg (30%) of the final product. LC-MS m/z 820 (M+H)⁺, 1.32 min (ret time).

Example 333 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({ethyl [2-(methylamino)ethyl]amino}methyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of 1,1-dimethylethyl 4-[(3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-1-piperazinecarboxylate (0.130 g, 0.16 mmol) in DCM (200 μL) was added TFA (100 μL, 0.1298 mmol) after which the mixture was stirred at room temperature for ¼ h. The reaction mixture was concentrated under vacuum, purified using a Gilson HPLC (with 0.1% TFA), and product concentrated using a GeneVac EZ-2 evaporator at 45° C. for 8 h. Extra TFA was removed by re-dissolving the resultant mixture in DCM and passing it though an amino cartridge to afford 0.0555 g (48%) of the title compound. LC-MS m/z 716 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.39 (t, J=7.53 Hz, 3H) 1.48 (t, J=7.28 Hz, 3H) 1.80-1.95 (m, 2H) 2.08 (d, J=12.30 Hz, 2H) 3.15 (q, J=7.53 Hz, 2H) 3.57 (s, 8H) 3.62-3.72 (m, 2H) 4.02 (dd, J=8.41, 3.14 Hz, 2H) 4.27-4.39 (m, 1H) 4.43-4.52 (m, 4H) 4.64 (s, 2H) 4.70 (s, 2H) 7.35-7.46 (m, 2H) 7.46-7.63 (m, 4H) 7.65 (s, 1H) 7.73 (d, J=7.53 Hz, 1H) 7.81 (s, 1H) 8.05 (d, J=7.78 Hz, 2H) 8.27 (s, 1H) 8.40 (s, 1H).

Example 334 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,4-benzenedicarboxamide

To a solution of 1,1-dimethylethyl 4-[(3′-{[({4-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)-amino]methyl}-3-biphenylyl)methyl]-1-piperazinecarboxylate (0.130 g, 0.16 mmol) in acetonitrile (7.2 mL) and H₂O (4800 μL) was added TFA (100 μL, 0.160 mmol). The resultant mixture was applied to a GeneVac EZ-2 evaporator at 45° C. for 8 h. The mixture was further purified using a Gilson HPLC (with 0.1% TFA) and the product-containing fraction concentrated on a GeneVav EZ-2 evaporator at 45° C. for 8 h. The extra TFA was removed by re-dissolving the resultant mixture in DCM and passing it though an amino cartridge. The mixture was concentrated down again by Glas-Col evaporator to afford the title compound 0.0600 g (52%). LC-MS m/z 716 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.40 (t, J=7.53 Hz, 3 H) 1.48 (t, J=7.15 Hz, 3 H) 1.81-1.97 (m, 2 H) 2.09 (d, J=12.05 Hz, 2 H) 3.16 (q, J=7.45 Hz, 2 H) 3.56 (s, 8 H) 3.68 (t, J=11.42 Hz, 2 H) 3.99-4.11 (m, J=8.41, 3.14 Hz, 2 H) 4.29-4.41 (m, J=4.27 Hz, 1 H) 4.43-4.53 (m, 4 H) 4.64 (s, 2 H) 4.71 (s, 2 H) 7.35-7.46 (m, 2 H) 7.47-7.61 (m, 3 H) 7.65 (s, 1 H) 7.74 (d, J=7.53 Hz, 1 H) 7.81 (s, 1 H) 7.92-8.03 (m, 4 H) 8.28 (s, 1 H).

Example 335 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of N-[(3-bromo-4-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.166 g, 0.26 mmol) in 1,4-dioxane (3 mL) and H₂O (1 mL) was added 1,1-dimethylethyl 4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperidinecarboxylate (0.157 g, 0.390 mmol), Pd(Ph₃P)₄ (0.015 g, 0.013 mmol), and K₂CO₃ (0.144 g, 1.040 mmol). This mixture was heated in a microwave at 130° C. for 15 min. The organic layer was separated, filtered, concentrated, and purified with a Gilson HPLC (with 0.1% TFA). Before the sample was concentrated with GeneVav EZ-2, TFA (5 drops) was added to each tube (12 mL). The resultant mixture was concentrated, purified with a Gilson HPLC (with 0.1% TFA), concentrated, and basified with an amino cartridge to afford 0.0738 g (39%) of the title compound. LC-MS m/z 732 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.36-1.52 (m, 8 H) 1.81-1.93 (m, 4 H) 2.09 (d, J=11.54 Hz, 2 H) 2.62-2.70 (m, 2 H) 2.87-2.98 (m, 2 H) 3.16 (q, 2 H) 3.19-3.30 (m, 1 H) 3.32-3.39 (m, J=12.55 Hz, 2 H) 3.62-3.71 (m, 2 H) 3.99-4.07 (m, 2 H) 4.29-4.40 (m, J=4.27, 4.27 Hz, 1 H) 4.48 (q, J=7.11 Hz, 2 H) 4.60 (s, 2 H) 4.68-4.75 (m, 2 H) 7.14 (dd, J=10.54, 8.53 Hz, 1 H) 7.18-7.26 (m, 1 H) 7.33-7.44 (m, 4 H) 7.44-7.50 (m, J=7.53, 2.26 Hz, 1 H) 7.59 (t, J=7.78 Hz, 1 H) 8.01-8.08 (m, 2 H) 8.28 (s, 1 H) 8.39 (s, 1 H).

Example 336 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-hydroxy-1-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (0.133 g, 0.2 mmol) in DCM (3 mL) was added 4-piperidinol (0.021 g, 0.210 mmol), NaBH(OAc)₃ (0.051 g, 0.240 mmol). This mixture was stirred at room temperature for 15 h at which time more 4-piperidinol (0.004 g, 0.04 mmol) was added and stirring continued for 2 h. Then more NaBH(OAc)₃ (0.025 g, 0.1 mmol) was added and the mixture was stirred for an additional 2 h. The reaction mixture was filtered, concentrated, and purified using a Gilson HPLC (with 0.1% TFA). The fraction was concentrated and basified with an amine cartridge to afford 0.1067 g of the title compound. LC-MS m/z 748 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.40 (t, J=7.65 Hz, 3 H) 1.49 (t, J=7.15 Hz, 3 H) 1.62-1.83 (m, 1 H) 1.83-2.01 (m, 4 H) 2.05-2.27 (m, 3 H) 3.08 (t, J=12.05 Hz, 1 H) 3.16 (q, J=7.36 Hz, 2 H) 3.32-3.41 (m, 2 H) 3.53 (d, J=12.30 Hz, 1 H) 3.62-3.72 (m, 2 H) 3.76-4.10 (m, 3 H) 4.31-4.51 (m, 5 H) 4.61 (s, 2 H) 4.72 (s, 2 H) 7.18 (dd, J=10.67, 8.41 Hz, 1 H) 7.38-7.46 (m, 1 H) 7.50-7.63 (m, 4 H) 7.65-7.76 (m, 2 H) 8.01-8.09 (m, 2 H) 8.29 (s, 1 H) 8.40 (t, J=1.63 Hz, 1 H).

Example 337 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-oxo-1-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-hydroxy-1-piperidinyl)methyl]-3-biphenyl-yl}methyl)-1,3-benzenedicarboxamide (85 mg, 0.114 mmol) in DCM (0.5 mL) was added Dess-Martin Periodinane (0.417 mL, 0.125 mmol). This mixture was stirred at room temperature for 40 min. The reaction was quenched with Na₂SO₃ (˜0.2 mL, 5%), concentrated and purified using a Gilson HPLC (with 0.1% TFA). The product fraction was concentrated and basified with an amine cartridge to afford 0.0121 g (14%) of the title compound. LC-MS m/z 746 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.31 (t, J=7.40 Hz, 3 H) 1.41 (t, J=7.15 Hz, 3 H) 1.64-1.83 (m, 4 H) 1.99-2.02 (m, J=1.25 Hz, 1 H) 2.04-2.07 (m, 1 H) 2.38 (t, J=6.02 Hz, 2 H) 2.59-2.78 (m, 4 H) 3.00 (q, J=7.45 Hz, 2 H) 3.60 (t, J=11.29 Hz, 2 H) 3.65-3.69 (m, 1 H) 3.71-3.78 (m, 1 H) 3.93-4.02 (m, J=11.54 Hz, 2 H) 4.10-4.20 (m, 1 H) 4.41 (q, J=7.19 Hz, 2 H) 4.58 (s, 2 H) 4.69 (s, 2 H) 7.09-7.18 (m, 1 H) 7.31-7.50 (m, 5 H) 7.51-7.59 (m, 2 H) 7.95-8.03 (m, 3 H) 8.31 (s, 1 H).

Example 338 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(4-piperidinyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of 1,1-dimethylethyl 4-[(3-bromophenyl)oxy]-1-piperidinecarboxylate (0.036 g, 0.100 mmol) in 1,4-dioxane (0.9 mL) and H₂O (300 μL) was added N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1,3-benzenedicarboxamide (0.067 g, 0.1 mmol), Pd(Ph₃P)₄ (4.62 mg, 4.00 μmol), and K₂CO₃ (0.028 g, 0.200 mmol). This mixture was heated at 130° C. for 15 min. The reaction mixture was concentrated (Glas-Col) and purified with CombiFlash chromatograph. To the resultant material was added DCM (300 μL) and TFA (100 μL, 1.298 mmol) which was then stirred at room temperature for 30 min. The reaction mixture was concentrated (Glas-Col), purified using a Gilson HPLC (with 0.1% TFA), concentrated on a GeneVac EZ-2 and neutralized with an amine cartridge to afford 0.0237 g (33.1%) of the title compound. LC-MS m/z 716 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.39 (t, J=6.78 Hz, 3 H) 1.48 (t, J=6.90 Hz, 3 H) 1.80-1.97 (m, 2 H) 1.99-2.24 (m, 5 H) 3.09-3.28 (m, 4 H) 3.41 (t, J=9.29 Hz, 2 H) 3.66 (t, J=11.04 Hz, 2 H) 4.02 (d, J=8.78 Hz, 2 H) 4.33 (br. s., 1 H) 4.42-4.90 (m, 8 H) 6.61-6.85 (m, 1 H) 6.94-7.17 (m, 1 H) 7.21 (br. s, 2 H) 7.31-7.66 (m, 5 H) 8.04 (d, J=7.53 Hz, 2 H) 8.27 (br. s., 1 H) 8.35-8.44 (m, 1 H).

Example 339 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.03 g, 0.066 mmol) in DCM (0.66 mL) was added 1,1-dimethylethyl 4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-1-piperidinecarboxylate (0.025 g, 0.066 mmol), HBTU (0.028 g, 0.073 mmol), and TEA (0.014 mL, 0.100 mmol). This mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated (Glas-Col), purified using a Gilson HPLC (with 0.1% TFA) and concentrated with GeneVav EZ-2 evaporator. To the resultant mixture was added DCM (0.1 mL) and TFA (0.033 mL, 0.428 mmol). The resultant mixture was left at room temperature for 30 min before being concentrated on Glas-Col evaporator, purified using a Gilson HPLC (with 0.1% TFA), concentrated with GeneVav EZ-2 and basified with an amine cartridge to afford 0.0245 g (51.7%) of the title compound. LC-MS m/z 714 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.35-1.45 (m, J=7.40, 7.40 Hz, 4 H) 1.44-1.53 (m, J=7.15, 7.15 Hz, 4 H) 1.80-1.96 (m, J=13.80 Hz, 5 H) 2.08 (d, J=12.30 Hz, 2 H) 2.66 (d, J=6.53 Hz, 2 H) 2.92 (t, J=12.55 Hz, 2 H) 3.15 (q, J=7.45 Hz, 2 H) 3.35 (d, J=14.31 Hz, 2 H) 3.65 (t, J=11.29 Hz, 2 H) 4.02 (d, J=11.04 Hz, 2 H) 4.27-4.39 (m, 1 H) 4.47 (q, J=7.19 Hz, 2 H) 4.64 (s, 2 H) 4.70 (s, 2 H) 7.16 (d, J=7.28 Hz, 1 H) 7.30-7.54 (m, 6 H) 7.55-7.65 (m, 2 H) 8.05 (d, J=7.78 Hz, 2 H) 8.27 (s, 1 H) 8.40 (s, 1 H).

Example 340 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of ({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)amine (0.0294 g, 0.10 mmol) in DCM (1 mL) was added 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.0452 g, 0.10 mmol), HBTU (0.0417 g, 0.11 mmol), and TEA (0.0303 mL, 0.20 mmol) and was stirred at room temperature for 17 h. The reaction was quenched with H₂O (1 drop), concentrated by Glas-Col evaporator and re-dissolved in MeOH/DMSO (0.5/0.1 mL). The resultant mixture was purified using a Gilson HPLC (with 0.1% TFA), concentrated by GeneVac EZ-2 evaporator and basified with an amine cartridge to afford the title compound 0.0432 g (59.34%). LC-MS m/z 728 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.37-1.44 (m, 3 H) 1.46-1.54 (m, J=7.28, 7.28 Hz, 5 H) 1.81-1.98 (m, 5 H) 2.05-2.13 (m, J=12.55 Hz, 2 H) 2.68 (d, J=6.53 Hz, 2 H) 2.78-2.86 (m, 3 H) 2.88-2.97 (m, 2 H) 3.15 (q, J=7.61 Hz, 2 H) 3.43-3.51 (m, J=12.30 Hz, 2 H) 3.67 (t, J=11.54 Hz, 2 H) 4.03 (d, J=11.80 Hz, 2 H) 4.30-4.41 (m, 1 H) 4.47 (q, J=7.19 Hz, 2 H) 4.65 (s, 2 H) 4.71 (s, 2 H) 7.17 (d, J=7.53 Hz, 1 H) 7.33-7.53 (m, 6 H) 7.59-7.66 (m, 2 H) 8.05 (dd, J=7.91, 1.63 Hz, 2 H) 8.29 (s, 1 H) 8.39 (s, 1 H).

Example 341 N-({3′-[(4-Cyano-1-piperidinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

To a solution of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (82 mg, 0.129 mmol) in DCM (2.58 mL) was added 4-piperidinecarbonitrile (14.21 mg, 0.129 mmol), NaBH(OAc)₃ (41.0 mg, 0.193 mmol). This mixture was stirred at room temperature for 19 h. The reaction mixture was washed with H₂O (1 mL), concentrated, purified using a Gilson HPLC (with 0.1% TFA), and basified with an amine cartridge to afford 0.0274 g (28%) of the title compound. LC-MS m/z 757 (M+H)⁺; ¹H NMR (400 MHz, MeOD) δ ppm 1.41 (t, J=7.53 Hz, 3 H) 1.49 (t, J=7.15 Hz, 3 H) 1.82-1.93 (m, J=13.05 Hz, 2 H) 1.96-2.38 (m, 6 H) 2.92-3.20 (m, 4 H) 3.22-3.29 (m, 1 H) 3.49-3.73 (m, 4 H) 4.00-4.09 (m, 2 H) 4.31-4.52 (m, 5 H) 4.62 (s, 2 H) 4.72 (s, 2 H) 7.20 (dd, J=10.54, 8.53 Hz, 1 H) 7.39-7.46 (m, 1 H) 7.50-7.65 (m, 4 H) 7.67-7.77 (m, 2 H) 8.05 (d, J=8.03 Hz, 2 H) 8.30 (s, 1 H) 8.39 (t, J=1.51 Hz, 1 H).

Example 342 N′-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[6-fluoro-3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-N-methyl-1,3-benzenedicarboxamide

To 1,1-dimethylethyl 4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]phenyl}carbonyl)(methyl)amino]-methyl}-2′-fluoro-3-biphenylyl)methyl]-1-piperazinecarboxylate in DCM (0.8 mL) was added TFA (0.40 mL) and the mixture stirred at room temperature for 15 h. The reaction was quenched with NaHCO₃ (sat. aq.), extracted with DCM (2×5 mL), concentrated, purified using a Gilson HPLC (with TFA), and the final product was basified with an amine cartridge to afford 0.077 g of the title compound. LC-MS m/z 747 (M+H)⁺; ¹H-NMR (400 MHz, CD₃OD) δ ppm 1.41 (t, J=7.65 Hz, 3 H) 1.49 (t, J=7.28 Hz, 3 H) 1.75-1.92 (m, 2 H) 2.01-2.13 (m, 2 H) 2.96 (s, 2 H) 3.06-3.11 (m, 1 H) 3.12-3.21 (m, 2 H) 3.34-3.43 (m, 4 H) 3.45-3.54 (m, J=3.51 Hz, 4 H) 3.61-3.73 (m, 2 H) 3.96-4.08 (m, 2 H) 4.27-4.40 (m, 3 H) 4.47 (q, J=7.19 Hz, 2 H) 4.55-4.63 (m, 1 H) 4.68-4.76 (m, 2 H) 4.81 (s, 1 H) 7.18-7.31 (m, 2 H) 7.43-7.76 (m, 7 H) 7.96-8.04 (m, 2 H) 8.25-8.33 (m, 1 H).

Example 343 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-N-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]benzamide

A cooled solution (0° C.) of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine, (80 mg, 0.26 mmol) in dichloromethane (10 mL) was treated with 3-(chlorosulfonyl)benzoic acid (30 mg, 0.36 mmol) and diisopropylethylamine (0.09 mL, 0.52 mmol) and the mixture was stirred at 0° C. for 20 min. The mixture was then treated with PyBOP (100 mg, 0.31 mmol), diisopropylethylamine (0.15 mL, 0.86 mmol) and 1,1-dimethylethyl (2S)-4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate (107 mg, 0.26 mmol) and stirred at ambient temperature for 16 h. The mixture was treated with water (40 mL) and extracted with dichloromethane (25 mL) and the organic phase was then concentrated to dryness and the intermediate Boc-protected material purified by mass-directed automated preparative HPLC. Product-containing fractions were passed though a 5 g SCX solid-phase extraction cartridge and the product eluted off with a 10% solution of ammonia in methanol, product containing fractions were concentrated to dryness. The residue was dissolved in methanol, treated with aqueous hydrochloric acid (2M, 2 mL) and stirred at ambient temperature for 16 h. The mixture was then dried (hydrophobic frit), evaporated to dryness and the product purified by mass-directed automated preparative HPLC. Product-containing fractions were passed though a 2 g SCX solid-phase extraction cartridge and the product eluted off with a 10% solution of ammonia in methanol, product-containing fractions were concentrated to dryness to afford 16 mg of the title compound. LC-MS m/z 783 (M+H)⁺, 0.92 min (ret time).

Example 344 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-[({[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}amino)sulfonyl]benzamide

To a solution of 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-amine (0.0195 g, 0.0641 mmol) in DCM (0.64 mL) was added 3-{[({3′-[(4-{[(1,1-dimethyl ethyl)oxy]carbonyl}-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)amino]sulfonyl}benzoic acid (0.0374 g, 0.0641 mmol), HBTU (0.0365 g, 0.0962 mmol), Et₃N (0.018 mL, 0.128 mmol). The result mixture was stirred at room temperature for 21 h before it was concentrated and purified using a Gilson HPLC (w/TFA). The HPLC fraction was concentrated on a GeneVac EZ-2 evaporator at 45° C. (de-Boc completed during evaporation), basified with an amine cartridge to afford 0.0161 g (33%) of the title compound. LC-MS m/z 769 (M+H)⁺; ¹H-NMR (400 MHz, MeOD) δ ppm 1.41 (t, J=7.30 Hz, 3 H) 1.49 (t, J=7.05 Hz, 3 H) 1.86 (q, J=9.90 Hz, 2 H) 2.09 (d, J=12.34 Hz, 2 H) 3.17 (q, J=7.39 Hz, 2 H) 3.46-3.59 (m, J=12.59 Hz, 8 H) 3.67 (t, J=11.33 Hz, 2 H) 4.04 (d, J=11.83 Hz, 2 H) 4.16 (s, 2 H) 4.30-4.39 (m, 1 H) 4.41 (s, 2 H) 4.48 (q, J=6.88 Hz, 2 H) 4.70 (s, 2 H) 7.04 (t, J=9.44 Hz, 1 H) 7.18-7.27 (m, 1 H) 7.34 (d, J=7.05 Hz, 1 H) 7.50-7.58 (m, 3 H) 7.60-7.68 (m, 2 H) 7.98 (d, J=7.55 Hz, 1 H) 8.08 (d, J=7.81 Hz, 1 H) 8.30 (d, J=13.60 Hz, 2 H).

Example 345 2-Chloro-5-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-4-fluoro-N-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]benzamide

345(a) 5-(Aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine (30.0 mg, 0.1 mmol) and 2-chloro-5-(chlorosulfonyl)-4-fluorobenzoic acid (0.1 mmol) was dissolved in DCM (3 mL), followed by the addition of Et₃N (10 mg, 0.1 mmol). The resultant solution was stirred overnight. The solution was then purified using a Gilson to give 2-chloro-5-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-4-fluorobenzoic acid.

345(b) The 2-chloro-5-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-4-fluorobenzoic acid, was dissolved in 3 mL of DCM with 1,1-dimethylethyl (2S)-4-{[5′-(aminomethyl)-2′-fluoro-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate (41.3 mg, 0.1 mmol), and then added in HOBt (1.0 eq, 14.0 mg), and EDC (1.0 eq, 19.0 mg). The resultant solution was stirred overnight. The solution was then purified using a Gilson to give 1,1-dimethylethyl (2S)-4-[(5′-{[({2-chloro-5-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-4-fluorophenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate.

345(c) The 1,1-dimethylethyl (2S)-4-[(5′-{[({2-chloro-5-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-4-fluorophenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate was dissolved in 2 mL of dioxane:MeOH (3:1). Three drops of HCl (concentrated) were added to the resultant solution and heated at 60° C. for 1 h. The solution was applied to the amine column, rinsed with 10 mL of dioxane:MeOH (3:1) to afford 63.0 mg (75.4%) the title compound. LC-MS m/z 837 (M+H)⁺, 1.29 min (ret time).

Example 346 5-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-N-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-2-(methyloxy)benzamide

The title compound was prepared according to the general procedure of Example 105 substituting for 5-(chlorosulfonyl)-2-(methyloxy)benzoic acid (0.1 mmol) for 2-chloro-5-(chlorosulfonyl)-4-fluorobenzoic acid to afford 50.0 mg of the title compound (61.5%). LC-MS m/z (M+H)⁺ m/z 813.3 (M+H), 1.26 (ret time).

Example 347 3-{4-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]phenyl}-N-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]propanamide

The title compound was prepared according to the general procedure of Example 105 using 3-[4-(chlorosulfonyl)phenyl]propanoic acid (0.1 mmol) for 2-chloro-5-(chlorosulfonyl)-4-fluorobenzoic acid to afford 59.0 mg of the title compound (72.7%). LC-MS m/z 811.4 (M+H)⁺, 1.22 min (ret time).

Example 348 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4b]pyridin-5-yl]methyl}-3-[(methyl{[3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}amino)sulfonyl]benzamide

To a solution of 3-{[[(3-bromophenyl)methyl](methyl)amino]sulfonyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}benzamide (120 mg, 0.179 mmol) in 1,4-dioxane (3 mL) and H₂O (1 mL) was added {3-[(4-{[(1,1-dimethylethyl)oxy]carbonyl}-1-piperazinyl)methyl]phenyl}boronic acid (57.4 mg g, 0.179 mmol), Pd(Ph₃P)₄ (10.35 mg, 8.96 μmol), K₂CO₃ (74.3 mg, 0.538 mmol) and was heated in microwave at 130° C. for 15 min. The organic layer was separated, filtered, concentrated, and purified using a Gilson HPLC (with 0.1% TFA). Before the sample was concentrated with a GeneVac EZ-2, TFA (5 drops) was added to each tube (12 mL). The resultant mixture was concentrated, purified using a Gilson HPLC (with 0.1% TFA), concentrated, and basified with amino cartridge to afford 0.0408 g (30%) of the title compound. LC-MS m/z 766 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.41 (t, J=7.03 Hz, 3 H) 1.49 (t, J=6.53 Hz, 3 H) 1.87 (q, J=11.04 Hz, 2 H) 2.10 (d, J=12.55 Hz, 2 H) 2.67 (s, 3 H) 3.18 (q, J=7.11 Hz, 2 H) 3.35-3.57 (m, 8 H) 3.67 (t, J=11.42 Hz, 2 H) 4.03 (d, J=11.54 Hz, 2 H) 4.23-4.42 (m, 5 H) 4.48 (q, J=7.03 Hz, 2 H) 4.75 (s, 2 H) 7.33 (d, J=7.53 Hz, 1 H) 7.40-7.63 (m, 5 H) 7.68 (d, J=7.28 Hz, 1 H) 7.77 (s, 2 H) 8.05 (d, J=5.77 Hz, 1 H) 8.23 (d, J=7.78 Hz, 1 H) 8.29 (s, 1 H) 8.39 (s, 1 H).

Example 349 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-({methyl[(3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]amino}sulfonyl)benzamide

To a solution of 3-{[[(3-bromophenyl)methyl](methyl)amino]sulfonyl}-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}benzamide (120 mg, 0.179 mmol) in 1,4-dioxane (3 mL) and H₂O (1 mL) was added {3-[((3R)-4-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-1-piperazinyl)methyl]phenyl}boronic acid (59.9 mg, 0.179 mmol), Pd(Ph₃P)₄ (10.35 mg, 8.96 μmol), and K₂CO₃ (74.3 mg, 0.538 mmol) and the reaction was heated in microwave at 130° C. for 15 min. The organic layer was separated, filtered, concentrated, and purified using a Gilson HPLC (with 0.1% TFA). Before the sample was concentrated with a GeneVac EZ-2, TFA (5 drops) was added to each tube (12 mL). The resultant mixture was concentrated, purified using a Gilson HPLC (with 0.1% TFA), concentrated, and basified with amino cartridge to afford 0.0429 g (30%) of the title compound. LC-MS m/z 779 (M+H)⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 1.35-1.45 (m, 6 H) 1.48 (t, J=7.28 Hz, 3 H) 1.82-1.96 (m, 2 H) 2.10 (d, J=12.55 Hz, 2 H) 2.66 (s, 3 H) 3.18 (q, J=7.61 Hz, 3 H) 3.33-3.39 (m, 1 H) 3.42-3.53 (m, 1 H) 3.60-3.79 (m, 6 H) 3.98-4.10 (m, J=8.41, 3.14 Hz, 2 H) 4.27 (s, 2 H) 4.31-4.40 (m, 1 H) 4.41-4.54 (m, 4 H) 4.75 (s, 2 H) 7.31-7.37 (m, 1 H) 7.40-7.65 (m, 5 H) 7.68-7.85 (m, 3 H) 8.05 (d, J=7.28 Hz, 1 H) 8.23 (d, J=7.78 Hz, 1 H) 8.28 (s, 1 H) 8.39 (s, 1 H).

Example 350 3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)methyl]-N-[(4′-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]benzamide

To a solution of 1,1-dimethylethyl (2S)-4-{[4-fluoro-3′-({[(3-formylphenyl)-carbonyl]amino}methyl)-3-biphenylyl]methyl}-2-methyl-1-piperazinecarboxylate (195 mg, 0.36 mmol) in dichloromethane (5 mL) was added 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine, and (110 mg, 0.36 mmol) and sodium triacetoxyborohydride (115 mg. 0.54 mmol) and the resulting solution was stirred at ambient temperature for 16 h and then treated with aqueous hydrochloric acid (2M, 2 mL) and stirred for a further 2 h. The mixture was treated with water (20 mL) and extracted with dichloromethane (25 mL). The organic phase was concentrated to dryness and the residue dissolved in methanol (5 mL), treated with aqueous hydrochloric acid (2M, 2 mL) and stirred at ambient temperature for 16 h. The mixture was then concentrated to dryness and the product purified by mass-directed automated preparative HPLC. Product-containing fractions were concentrated to dryness to afford 25 mg of the title compound. LC-MS m/z 733 (M+H)⁺, 0.79 min (ret time).

Example 351 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-{[({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)amino]sulfonyl}benzamide

To a solution of 3-{[({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)amino]sulfonyl}benzoic acid (0.086 g, 0.102 mmol) in THF (1 mL) was added 5-(aminomethyl)-1,6-diethyl-N-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridin-4-amine hydrochloride (0.042 g, 0.122 mmol), HBTU (0.048 g, 0.133 mmol), and TEA (0.036 mL, 0.255 mmol) and the reaction was stirred at room temperature for 18 h. The reaction mixture was concentrated on Glas-Col evaporator, purified using a Gilson HPLC (with 0.1% TFA), concentrated, basified with amino cartridge to afford 0.0458 g (59%) of the title compound. LC-MS m/z 764 (M+H)⁺.

Example 352 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

To a solution of 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (0.045 g, 0.1 mmol) in DMSO (0.5 mL) was added 1-{3′-[(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)methyl]-3-biphenylyl}methanamine dihydrochloride (0.047 g, 0.12 mmol), HBTU (0.054 g, 0.15 mmol), and TEA (0.042 mL, 0.3 mmol) and the reaction was stirred at room temperature for 16 h. The reaction mixture was purified using a Gilson HPLC (with 0.1% TFA), concentrated, basified with amino cartridge to afford 0.0409 g (52%) of the title compound. LC-MS m/z 754 (M+H)⁺.

Example 353 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[2-(3-{[(3S)-3-methyl-1-piperazinyl]methyl}phenyl)-4-pyridinyl]methyl}-1,3-benzenedicarboxamide

To 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (30 mg, 0.066 mmol) was added 1,1-dimethylethyl (2S)-4-({3-[4-(aminomethyl)-2-pyridinyl]phenyl}methyl)-2-methyl-1-piperazinecarboxylate (26.3 mg, 0.066 mmol) followed by HBTU (30.2 mg, 0.080 mmol) and TEA (18.52 μl, 0.133 mmol) in dichloromethane (DCM) (664 μl) to give a reaction mixture that was stirred at room temperature overnight. Then the reaction was quenched by H₂O (1 drop) and the solvent was removed by Glas-Col. The crude product was redissolved in MeOH/DMSO and purified using a Gilson HPLC (acidic conditions), concentrated by GeneVav EZ-2 evaporator. To this product was added TFA (102 μl, 1.329 mmol) in dichloromethane (0.2 mL) then stirred at room temperature overnight. The reaction mixture was dissolved in MeOH/DMSO and purified using a Gilson HPLC (acidic conditions), concentrated by GeneVav EZ-2 evaporator and basified with an amine cartridge to afford 16.6 mg (33%) of the title compound. LC-MS m/z 730 (M+H)⁺.

Example 354 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-[3-(4-piperidinylmethyl)phenyl]-4-pyridinyl}methyl)-1,3-benzenedicarboxamide

To 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]benzoic acid (30 mg, 0.066 mmol) was added 1,1-dimethylethyl 4-({3-[4-(aminomethyl)-2-pyridinyl]phenyl}methyl)-1-piperidinecarboxylate (25.3 mg, 0.066 mmol) followed by HBTU (30.2 mg, 0.080 mmol) and TEA (13.45 mg, 0.133 mmol) in dichloromethane (DCM) (664 μl) to give a reaction mixture that was stirred at room temperature overnight. Then the reaction was quenched with H₂O (1 drop) and the solvent was removed by Glas-Col. The crude product was redissolved in MeOH/DMSO and purified using a Gilson HPLC (acidic conditions) and concentrated by GeneVav EZ-2 evaporator. To this product was added TFA (93 μl, 1.329 mmol) in dichloromethane (0.2 mL) which was then stirred at room temperature overnight. The reaction mixture was dissolved in MeOH/DMSO and purified using a Gilson HPLC (acidic conditions), concentrated by GeneVav EZ-2 evaporator and basified with an amine cartridge to afford 17.1 mg (32%) of the title compound. LC-MS m/z 715 (M+H)⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 1.23-1.42 (m, 6 H) 1.68-1.76 (m, 3 H) 1.79-2.02 (m, 3 H) 2.08 (s, 2 H) 2.63 (d, J=6.78 Hz, 2 H) 2.76-2.84 (m, 2 H) 3.07-3.15 (m, 2 H) 3.20-3.28 (m, 2 H) 3.32-3.39 (m, 1 H) 3.58-3.66 (m, 2 H) 3.90 (dd, J=8.03, 3.51 Hz, 2 H) 4.29-4.37 (m, 1 H) 4.47 (q, J=7.19 Hz, 2 H) 4.60 (t, J=6.40 Hz, 4 H) 7.28 (d, J=7.78 Hz, 1 H) 7.34 (d, J=3.76 Hz, 1 H) 7.39-7.46 (m, 1 H) 7.65 (t, J=7.78 Hz, 1 H) 7.85 (d, J=8.03 Hz, 1 H) 7.92 (d, J=14.56 Hz, 2 H) 8.05 (d, J=8.03 Hz, 1 H) 8.12 (d, J=7.78 Hz, 1 H) 8.26 (br. s., 1 H) 8.40 (s, 2 H) 8.61 (d, J=4.52 Hz, 1 H) 8.86 (br. s., 1 H) 9.32-9.39 (m, 2 H).

Example 355 N-[(3′-{[(1S,4S)-5-Acetyl-2,5-diazabicyclo [2.2.2]oct-2-yl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (50.0 mg, 0.076 mmol) was diluted in DMSO (1.0 mL) and dispensed into a 1 dram vial, with fitted magnetic stir bar, containing (1S,4S)-2,5-diazabicyclo[2.2.2]octane (0.76 mmol), ZnCl₂ (0.032 mmol, 4.2 mg), NaB(OAc)₃H (0.76 mmol, 180 mg) for 12 h. Purification was performed via Gilson HPLC (with 0.1% TFA in the solvents) and the desired product was collected. The desired product was dissolved in dichloromethane (1 mL) and washed through 2 g SPE-Amine column (EtOAc:MeOH=4:1, 5 mL) to afford 7.96 mg (13.1%). of the title compound. LC-MS m/z 803 (M+H)⁺, 1.4 min (ret time).

Example 356 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (30 mg, 0.044 mmol), 1,1-dimethylethyl (2S)-2-methyl-1-piperazinecarboxylate (89 mg, 0.44 mmol, 10 eq) and acetic acid (2.54 μL, 0.044 mmol, 1 eq) were dissolved in DMSO (1.5 mL). The mixture was stirred in VX-2500 Multi-Tube Vortexer for overnight at room temperature. MP-triacetoxyborohydride (133 mg, 0.310 mmol, 7 eq) was then added and the mixture was stirred again in VX-2500 Multi-Tube Vortexer for overnight at room temperature. The reaction mixture was filtered through a polypropylene cartridge (10 mL tube) on Bohdan Miniblock in a reaction tube and concentrated in a Glas-Col evaporator. Methanol (2 mL) then hydrochloric acid (5 μL) were added for the deprotection of 1,1-dimethylethyl (2S)-4-[(5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylphenyl}carbonyl)amino]methyl}-2′-fluoro-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate. The vial containing the reaction mixture was closed and stirred in a Glas-Col evaporator for overnight at 60° C. The reaction mixture was then concentrated and purified using a Gilson HPLC with a water-acetonitrile with 0.1% TFA buffer. The desired product fractions were combined, filtered through amine cartouche (500 mg) on Bohdan Miniblock and concentrated in a Glas-Col evaporator, giving 20.16 mg (53.8%) of the title compound. LC-MS m/z 761 (M+H)⁺, 1.31 min (ret time).

Examples 357-362

Using array chemistry, following the procedure as described for the preparation of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (Example 356), N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide was reacted with an appropriate amine to give Examples 357-362 listed in Table 1.

TABLE 1 Examples 357-362. LC-MS m/z RT Example R Name (M + H)⁺ (min) 357

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran- 4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-{[6-fluoro-3′-(1- piperazinylmethyl)-3- biphenylyl]methyl}-5-methyl-1,3- benzenedicarboxamide 747 1.27 358

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran- 4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-[(6-fluoro-3′-{[(3R)-3- methyl-1-piperazinyl]methyl}-3- biphenylyl)methyl]-5-methyl-1,3- benzenedicarboxamide 761 1.31 359

N-({3′-[(1S,4S)-2,5- diazabicyclo[2.2.1]hept-2-ylmethyl]-6- fluoro-3-biphenylyl}methyl)-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-1,3- benzenedicarboxamide 759 1.26 360

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran- 4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-{[6-fluoro-3′-(hexahydro- 1H-1,4-diazepin-1-ylmethyl)-3- biphenylyl]methyl}-5-methyl-1,3- benzenedicarboxamide 761 1.25 361

N-[(3′-{[(3S)-3-amino-1- pyrrolidinyl]methyl}-6-fluoro-3- biphenylyl)methyl]-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo [3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 747 1.29 362

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran- 4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-[(6-fluoro-3′-{[(2S)-2- methyl-1-piperazinyl]methyl}-3- biphenylyl)methyl]-5-methyl-1,3- benzenedicarboxamide 761 0.74

Example 363 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (30 mg, 0.046 mmol), 1,1-dimethylethyl (2S)-2-methyl-1-piperazinecarboxylate (91 mg, 0.46 mmol, 10 eq) and acetic acid (2.61 μL, 0.046 mmol, 1 eq) were dissolved in DMSO (1.5 mL). The mixture was stirred in a VX-2500 Multi-Tube Vortexer overnight at room temperature. MP-triacetoxyborohydride (137 mg, 0.319 mmol, 7 eq) was then added and the mixture was stirred again in the VX-2500 Multi-Tube Vortexer overnight at room temperature. The reaction mixture was filtered through a polypropylene cartridge (10 mL tube) on Bohdan Miniblock (Artisian Scientific, Chanpaign, Ill., USA; http://www.artisan-scientific.com/51413.htm) in a reaction tube and concentrated in a Glas-Col evaporator. Methanol (2 mL), then hydrochloric acid (5 μL), were added for the deprotection of 1,1-dimethylethyl (2S)-4-[(3′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylphenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-2-methyl-1-piperazinecarboxylate. The vial containing the reaction mixture was closed and stirred in a Glas-Col evaporator for overnight at 60° C. The reaction mixture was then concentrated and purified using a Gilson HPLC with a water-acetonitrile-with-0.1% TFA buffer. The desired product fractions were combined, filtered through amine cartridge (500 mg) on a Bohdan Miniblock and concentrated in a Glas-Col evaporator, giving 22.99 mg (61.2%) of the title compound. LC-MS m/z 743.4 (M+H)⁺, 1.27 min (ret time); ¹H NMR (500 MHz, DMSO-d₆) δ 0.86 (d, J=6.35 Hz, 3 H), 1.24 (t, J=7.32 Hz, 3 H), 1.33 (t, J=7.08 Hz, 3 H), 1.52-1.58 (m, 2 H), 1.83-1.97 (m, 2 H), 2.40 (s, 3 H), 2.59-2.65 (m, 4 H), 2.70-2.76 (m, 1 H), 2.96 (q, J=7.49 Hz, 2 H), 3.16 (d, J=5.37 Hz, 3 H), 3.54 (t, J=10.50 Hz, 2 H), 3.83-3.89 (m, 2 H), 4.09 (q, J=5.37 Hz, 2 H), 4.32 (q, J=7.16 Hz, 2 H), 4.54 (d, J=5.86 Hz, 4 H), 7.05 (d, J=7.81 Hz, 1 H), 7.26 (d, J=7.32 Hz, 1 H), 7.31 (d, J=7.81 Hz, 1 H), 7.40 (ddd, J=12.09, 7.69, 7.57 Hz, 2 H), 7.47-7.53 (m, 3 H), 7.58 (s, 1 H), 7.80 (s, 1 H), 7.85 (s, 1 H), 8.00 (s, 1 H), 8.14 (s, 1 H), 8.97 (t, J=5.86 Hz, 1 H), 9.11 (t, J=5.86 Hz, 1 H).

Examples 364-367

Using array chemistry, following the procedure as described for the preparation of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (Example 363), N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide was reacted with an appropriate amine to give Examples 364-367 listed in Table 2.

TABLE 2 Examples 364-367. LC-MS m/z RT Example R Name (M + H)⁺ (min) 364

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl-N′- {[3′-(1-piperazinylmethyl)-3- biphenylyl]methyl}-1,3- benzenedicarboxamide 729.4 1.25 365

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl-N′- [(3′-{[(3R)-3-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-1,3- benzenedicarboxamide 743.4 1.26 366

N-({3′-[(1S,4S)-2,5- diazabicyclo[2.2.1]hept-2-ylmethyl]- 3-biphenylyl}methyl)-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin- 5-yl]methyl}-5-methyl-1,3- benzenedicarboxamide 741.4 1.21 367

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl-N′- [(3′-{[(2S)-2-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-1,3- benzenedicarboxamide 743.4 1.28

Example 368 N-{[6-Chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (30.36 mg, 0.044 mmol), 1,1-dimethylethyl 1-piperazinecarboxylate (82 mg, 0.44 mmol, 10 eq) and acetic acid (2.51 μL, 0.044 mmol, 1 eq) were dissolved in DMSO (1.5 mL). The mixture was stirred in VX-2500 Multi-Tube Vortexer for overnight at room temperature. MP-triacetoxyborohydride (133 mg, 0.310 mmol, 7 eq) was then added and the mixture was stirred again in VX-2500 Multi-Tube Vortexer for overnight at room temperature. The reaction mixture was filtered through a polypropylene cartridge (10 mL tube) on Bohdan Miniblock in a reaction tube and concentrated in a Glas-Col evaporator. Methanol (2 mL) then hydrochloric acid (5 μL) were added for the deprotection of 1,1-dimethylethyl 4-[(2′-chloro-5′-{[({3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylphenyl}carbonyl)amino]methyl}-3-biphenylyl)methyl]-1-piperazinecarboxylate. The vial containing the reaction mixture was closed and stirred in a Glas-Col evaporator for overnight at 60° C. The reaction mixture was then concentrated and purified using a Gilson HPLC with a water-acetonitrile with 0.1% TFA buffer. The desired product fractions were combined, filtered through amine cartouche (500 mg) on Bohdan Miniblock and concentrated in a Glas-Col evaporator, giving 23.82 mg (64.1%) of the title compound. LC-MS m/z 763 (M+H)⁺, 1.32 min (ret time).

Examples 369-374

Using array chemistry, following the procedure as described for the preparation of N-{[6-chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (Example 368), N-[(6-chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide was reacted with an appropriate amine to give Examples 369-374 listed in Table 3.

TABLE 3 Examples 369-374. LC-MS m/z RT Example R Name (M + H)⁺ (min) 369

N-[(6-chloro-3′-{[(3R)-3-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 777 1.78 370

N-({6-chloro-3′-[(1S,4S)-2,5- diazabicyclo[2.2.1]hept-2-ylmethyl]-3- biphenylyl}methyl)-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 775 1.28 371

N-[(6-chloro-3′-{[(3S)-3-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 777 1.34 372

N-{[6-chloro-3′-(hexahydro-1H-1,4- diazepin-1-ylmethyl)-3- biphenylyl]methyl}-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 777 1.27 373

N-[(3′-{[(3S)-3-amino-1- pyrrolidinyl]methyl}-6-chloro-3- biphenylyl)methyl]-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 763 1.38 374

N-[(6-chloro-3{[(2S)-2-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 777 1.36

Example 375 N-({6-Chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide

N-[(6-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (30.36 mg, 0.044 mmol), 1-methylpiperazine (43.9 mg, 0.44 mmol, 10 eq) and acetic acid (2.51 μL, 0.044 mmol, 1 eq) were dissolved in DMSO (1.5 mL). The mixture was stirred in VX-2500 Multi-Tube Vortexer for overnight at room temperature. MP-triacetoxyborohydride (133 mg, 0.310 mmol, 7 eq) was then added and the mixture was stirred again in VX-2500 Multi-Tube Vortexer for overnight at room temperature. The reaction mixture was filtered through a polypropylene cartridge (10 mL tube) on Bohdan Miniblock in a reaction tube and concentrated in a Glas-Col evaporator. The reaction mixture was then purified using a Gilson HPLC with a water-acetonitrile with 0.1% TFA buffer. The desired product fractions were combined, filtered through amine cartouche (500 mg) on Bohdan Miniblock and concentrated in a Glas-Col evaporator, giving 18.38 mg (48.6%) of the title compound. LC-MS m/z 777 (M+H)⁺, 1.41 min (ret time).

Examples 376-381

Using array chemistry, following the procedure as described above in the preparation of N-({6-chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (Example 375), N-[(6-chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide was reacted with an appropriate amine to give Examples 376-381 listed in Table 4.

TABLE 4 Examples 376-381 LC-MS m/z RT Example R Name (M + H)⁺ (min) 376

N-({6-chloro-3[(4-methylhexahydro-1H- 1,4-diazepin-1-yl)methyl]-3- biphenylyl}methyl)-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 791 1.32 377

N-({3′-[(4-acetyl-1-piperazinyl)methyl]-6- chloro-3-biphenylyl}methyl)-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4-ylamino)- 1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 805 1.43 378

N-({6-chloro-3′-[(4-ethyl-1- piperazinyl)methyl]-3-biphenylyl}methyl)- {[1,6-diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-1,3- benzenedicarboxamide 791 1.42 379

N-[(6-chloro-3′-{[[2- (dimethylamino)ethyl](methyl)amino]methyl}- 3-biphenylyl)methyl]-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 779 1.32 380

N-{[6-chloro-3′-(1-piperidinylmethyl)-3- biphenylyl]methyl}-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 762 1.53 381

N-[(6-chloro-3′-{[(3R,5S)-3,5-dimethyl-1- piperazinyl]methyl}-3-biphenylyl)methyl]- N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-1,3- benzenedicarboxamide 791 1.39

Example 382 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-5-methyl-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (30 mg, 0.044 mmol), 1-methylpiperazine (44.4 mg, 0.44 mmol, 10 eq) and acetic acid (2.54 μL, 0.044 mmol, 1 eq) were dissolved in DMSO (1.5 mL). The mixture was stirred in VX-2500 Multi-Tube Vortexer for overnight at room temperature. MP-triacetoxyborohydride (133 mg, 0.310 mmol, 7 eq) was then added and the mixture was stirred again in VX-2500 Multi-Tube Vortexer for overnight at room temperature. The reaction mixture was filtered through a polypropylene cartridge (10 mL tube) on Bohdan Miniblock in a reaction tube and concentrated in a Glas-Col evaporator. The reaction mixture was then purified using a Gilson HPLC with a water-acetonitrile with 0.1% TFA buffer. The desired product fractions were combined, filtered through amine cartouche (500 mg) on Bohdan Miniblock and concentrated in a Glas-Col evaporator, giving 22.15 mg (59.1%) of the title compound. LC-MS m/z 761 (M+H)⁺, 1.38 min (ret time).

Examples 383-388

Using array chemistry, following the procedure as described above in the preparation of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-5-methyl-1,3-benzenedicarboxamide (Example 382), N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide was reacted with an appropriate amine to give Examples 383-388 listed in Table 5.

TABLE 5 Examples 383-388 LC-MS m/z RT Example R Name (M + H)⁺ (min) 383

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-N′-({6-fluoro- 3′-[(4-methylhexahydro-1H-1,4- diazepin-1-yl)methyl]-3- biphenylyl}methyl)-5-methyl-1,3- benzenedicarboxamide 775 1.29 384

N-({3′-[(4-acetyl-1- piperazinyl)methyl]-6-fluoro-3- biphenylyl}methyl)-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 789 1.37 385

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-N′-({3′-[(4- ethyl-1-piperazinyl)methyl]-6-fluoro-3- biphenylyl}methyl)-5-methyl-1,3- benzenedicarboxamide 775 1.32 386

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-N′-[(3′-{[[2- (dimethylamino)ethyl](methyl)amino] methyl}-6-fluoro-3-biphenylyl)methyl]-5- methyl-1,3-benzenedicarboxamide 763 1.27 387

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-N′-{[6-fluoro- 3′-(1-piperidinylmethyl)-3- biphenylyl]methyl}-5-methyl-1,3- benzenedicarboxamide 746 1.43 388

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-N′-[(3′- {[(3R,5S)-3,5-dimethyl-1- piperazinyl]methyl}-6-fluoro-3- biphenylyl)methyl]-5-methyl-1,3- benzenedicarboxamide 775 1.38

Example 389 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide (30 mg, 0.046 mmol), 1-methylhexahydro-1H-1,4-diazepine (52 mg, 0.46 mmol, 10 eq) and acetic acid (2.61 μL, 0.046 mmol, 1 eq) were dissolved in DMSO (1.5 mL). The mixture was stirred in VX-2500 Multi-Tube Vortexer for overnight at room temperature. MP-triacetoxyborohydride (137 mg, 0.319 mmol, 7 eq) was then added and the mixture was stirred again in VX-2500 Multi-Tube Vortexer for overnight at room temperature. The reaction mixture was filtered through a polypropylene cartridge (10 mL tube) on Bohdan Miniblock in a reaction tube and concentrated in a Glas-Col evaporator. The reaction mixture was then purified using a Gilson HPLC with a water-acetonitrile with 0.1% TFA buffer. The desired product fractions were combined, filtered through amine cartouche (500 mg) on Bohdan Miniblock and concentrated in a Glas-Col evaporator, giving 9.74 mg (25.4%) of the title compound. LC-MS m/z 757.4 (M+H)⁺, 1.26 min (ret time).

Examples 390-394

Using array chemistry, following the procedure as described above in the preparation of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide (Example 389), N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-formyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide was reacted with an appropriate amine to give Examples 390-394 listed in Table 6.

TABLE 6 Examples 390-394. LC-MS m/z RT Example R Name (M + H)⁺ (min) 390

N-({3[(4-acetyl-1-piperazinyl)methyl]-3- biphenylyl}methyl)-N′-{[1,6-diethyl-4- (tetrahydro-2H-pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5-yl]methyl}-5- methyl-1,3-benzenedicarboxamide 771.4 1.35 391

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-({3′-[(4-ethyl-1- piperazinyl)methyl]-3-biphenylyl}methyl)- 5-methyl-1,3-benzenedicarboxamide 757.4 1.35 392

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-[(3′-{[[2- (dimethylamino)ethyl](methyl)amino]methyl}- 3-biphenylyl)methyl]-5-methyl-1,3- benzenedicarboxamide 745.4 1.24 393

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-{[3′-(1- piperidinylmethyl)-3-biphenylyl]methyl}- 1,3-benzenedicarboxamide 728.4 1.44 394

N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl- 1-piperazinyl]methyl}-3- biphenylyl)methyl]-5-methyl-1,3- benzenedicarboxamide 757.4 1.30

Example 395 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide (40.1 mg. 0.06 mmol) was diluted in DMSO (1.5 mL) and dispensed into a 1 dram vial with fitted magnetic stir bar containing 1,1-dimethylethyl (2S)-2-methyl-1-piperazinecarboxylate (0.18 mmol), acetic acid (3.6 mg, 0.6 mmol). The resulting solution was stirred at room temperature for 4 h. MP-B(OAc)₃H (0.6 mmol, 140 mg) was added and the solution was stirring for another 12 h. The polymer reagent was filtered and to the solution added MeOH (2.0 mL) and 1 drop of concentrated HCl. The solution was heated at 60° C. for 12 h. Purification was completed via Gilson HPLC (with 0.1% TFA in the solvents). The product was dissolved in 3 mL of MeOH and passed through 0.5 g amine columns (washed with 8 mL MeOH) to afford 24.66 mg (53.16%) of the title compound. LC-MS m/z 773 (M+H)⁺, 1.253 min (ret time).

Examples 396-405

Using array chemistry, following the procedure as described above in the preparation of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (Example 395), an appropriate aldehyde was reacted with an appropriate amine to give Examples 396-405 listed in Table 7.

TABLE 7 Examples 396-405. LC-MS m/z RT Example R1 R2 Name (M + H)⁺ (min) 396 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-[(6- (methyloxy)-3′-{[(3R)-3-methyl- 1-piperazinyl]methyl}-3- biphenylyl)methyl]-1,3- benzenedicarboxamide 774 1.235 397 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-[(6- methyl-3′-{[(3R)-3-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-1,3- benzenedicarboxamide 758 1.276 398 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-[(6- methyl-3-{[(3S)-3-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-1,3- benzenedicarboxamide 758 1.281 399 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-{[3′-(hexahydro- 1H-1,4-diazepin-1-ylmethyl)-6- (methyloxy)-3- biphenylyl]methyl}-5-methyl-1,3- benzenedicarboxamide 774 1.173 400 OMe

N-{[3′-[(1R,4S)-2,5- diazabicyclo[2.2.1]hept-2- ylmethyl]-6-(methyloxy)-3- biphenylyl]methyl}-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl- 1,3-benzenedicarboxamide 772 1.249 401 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-[(6- (methyloxy)-3′-{[(2S)-2-methyl- 1-piperazinyl]methyl}-3- biphenylyl)methyl]-1,3- benzenedicarboxamide 774 1.275 402 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-{[3′-(hexahydro- 1H-1,4-diazepin-1-ylmethyl)-6- methyl-3-biphenylyl]methyl}-5- methyl-1,3- benzenedicarboxamide 758 1.223 403 Me

N-[(3′-{[(3S)-3-amino-1- pyrrolidinyl]methyl}-6-methyl-3- biphenylyl)methyl]-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl- 1,3-benzenedicarboxamide 744 1.191 404 Me

N-{[3′-(2,5- diazabicyclo[2.2.1]hept-2- ylmethyl)-6-methyl-3- biphenylyl]methyl}-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl- 1,3-benzenedicarboxamide 755 1.239 405 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-[(6- methyl-3′-{[(2S)-2-methyl-1- piperazinyl]methyl}-3- biphenylyl)methyl]-1,3- benzenedicarboxamide 758 1.303

Example 406 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[6-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-formyl-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide (40.1 mg. 0.06 mmol) was diluted in DMSO (1.5 mL) and dispensed into a 1 dram vial, with fitted magnetic stir bar, containing piperazine (0.18 mmol), and acetic acid (3.6 mg, 0.6 mmol). The resulting solution was stirred at room temperature for 4 h. MP-B(OAc)₃H (0.6 mmol, 140 mg) was added and the solution was stirred for another 12 h. The polymer reagent was filtered and purification was completed via Gilson HPLC (with 0.1% TFA in the solvents). The product was dissolved in 3 mL of MeOH and passed through 0.5 g amine columns (washed with 8 mL MeOH) to afford 29.8 mg (65.3%) of the title compound. LC-MS m/z 760 (M+H)⁺, 1.224 min (ret time).

Examples 407-421

Using array chemistry, following the procedure as described above in the preparation of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[6-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (Example 406), an appropriate aldehyde was reacted with an appropriate amine to give Examples 407-421 listed in Table 8.

TABLE 8 Examples 407-421. LC-MS m/z RT Example R1 R2 Name (M + H)⁺ (min) 407 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-{[6- methyl-3′-(1-piperazinylmethyl)- 3-biphenylyl]methyl}-1,3- benzenedicarboxamide 744 1.282 408 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-({6- (methyloxy)-3′-[(4-methyl-1- piperazinyl)methyl]-3- biphenylyl}methyl)-1,3- benzenedicarboxamide 773 1.283 409 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-({6- methyl-3′-[(4-methyl-1- piperazinyl)methyl]-3- biphenylyl}methyl)-1,3- benzenedicarboxamide 757 1.342 410 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-{[3′-[(4- methylhexahydro-1H-1,4- diazepin-1-yl)methyl]-6- (methyloxy)-3- biphenylyl]methyl}-1,3- benzenedicarboxamide 787 1.182 411 OMe

N-{[3′-[(4-acetyl-1- piperazinyl)methyl]-6- (methyloxy)-3- biphenylyl]methyl}-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl- 1,3-benzenedicarboxamide 801 1.443 412 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-{[3′-[(4-ethyl-1- piperazinyl)methyl]-6- (methyloxy)-3- biphenylyl]methyl}-5-methyl-1,3- benzenedicarboxamide 787 1.283 413 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-{[3′-{[[2- (dimethylamino)ethyl](methyl) amino]methyl}-6-(methyloxy)-3- biphenylyl]methyl}-5-methyl-1,3- benzenedicarboxamide 775 1.183 414 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-{[6- (methyloxy)-3′-(1- piperidinylmethyl)-3- biphenylyl]methyl}-1,3- benzenedicarboxamide 758 1.398 415 OMe

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-{[3′-{[(3R,5S)-3,5- dimethyl-1-piperazinyl]methyl}- 6-(methyloxy)-3- biphenylyl]methyl}-5-methyl-1,3- benzenedicarboxamide 787 1.249 416 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-({6- methyl-3′-[(4-methylhexahydro- 1H-1,4-diazepin-1-yl)methyl]-3- biphenylyl}methyl)-1,3- benzenedicarboxamide 771 1.227 417 Me

N-({3′-[(4-acetyl-1- piperazinyl)methyl]-6-methyl-3- biphenylyl}methyl)-N′-{[1,6- diethyl-4-(tetrahydro-2H-pyran-4- ylamino)-1H-pyrazolo[3,4- b]pyridin-5-yl]methyl}-5-methyl- 1,3-benzenedicarboxamide 785 1.369 418 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-({3′-[(4-ethyl-1- piperazinyl)methyl]-6-methyl-3- biphenylyl}methyl)-5-methyl-1,3- benzenedicarboxamide 771 1.348 419 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-[(3′-{[[2- (dimethylamino)ethyl](methyl) amino]methyl}-6-methyl-3- biphenylyl)methyl]-5-methyl-1,3- benzenedicarboxamide 759 1.195 420 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-5-methyl-N′-{[6- methyl-3′-(1-piperidinylmethyl)- 3-biphenylyl]methyl}-1,3- benzenedicarboxamide 742 1.464 421 Me

N-{[1,6-diethyl-4-(tetrahydro-2H- pyran-4-ylamino)-1H- pyrazolo[3,4-b]pyridin-5- yl]methyl}-N′-[(3′-{[(3R,5S)-3,5- dimethyl-1-piperazinyl]methyl}- 6-methyl-3-biphenylyl)methyl]-5- methyl-1,3- benzenedicarboxamide 771 1.35

Example 422 N-({5-Chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(5-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (34.0 mg, 0.05 mmol), 1-methylpiperazine (10.12 mg, 0.100 mmol), MP-Triacetoxyborohydride (107 mg, 0.250 mmol), and acetic acid (3.15 μL, 0.055 mmol) were combined in dimethyl sulfoxide (DMSO) (0.5 mL) in a 1 dram vial and stirred at room temperature overnight. The mixture was filtered through glass fiber filter paper and washed 2× with 0.3 mL of DMSO. The combined filtrates were purified on a Gilson HPLC eluting at 20 mL/min with a linear gradient running from 0% CH₃CN/H₂O (0.1% TFA) to 90% CH₃CN/H₂O (0.1% TFA). The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated, the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-({5-chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide as a white solid (18.4 mg, 47%). LC-MS m/z 763 M⁺; mp 116-118° C.

Example 423 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide

In an alternate process for the preparation of the title compound, N-{[3-bromo-4-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (40 mg, 0.060 mmol), 1,1-dimethylethyl (2S)-2-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperazinecarboxylate (25.10 mg, 0.060 mmol), potassium carbonate (24.99 mg, 0.181 mmol), and Pd(Ph₃P)₄ (3.48 mg, 3.01 μmol) were added to a 0.5-2 mL Biotage microwave vial in 1,4-dioxane (1.5 mL) and water (0.5 mL). The vial was capped and the mixture was heated under microwave at normal power in the Emrys Optimizer at 100° C. for 15 min. The crude product was partitioned between EtOAc (30 mL) and water (10 mL). The phases were separated, and the organic phase washed with brine, dried over anhydrous Na₂SO₄ filtered and concentrated to give a crude intermediate. It was taken up in DCM (1.8 mL) and treated with trifluoroacetic acid (0.2 mL). The mixture was stirred under argon for 5 hours. Solvents were pumped off and the residue taken up in DMSO and purified on the Gilson with the TFA system. Evaporation of the appropriate purified fractions gave the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated, the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide as a white solid (30.07 mg, 63.3%). LC-MS m/z 773 (M+H)⁺; mp 121-123° C.

Example 424 N-{[5-Chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(5-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (40 mg, 0.059 mmol) and 1,1-dimethylethyl 1-piperazinecarboxylate (21.94 mg, 0.118 mmol) were dissolved in 1,2-dichloroethane (DCE) (1 mL) in a 1 dram vial and a acetic acid (3.71 μL, 0.065 mmol) was added. The mixture was stirred for 30 min and then the MP-Triacetoxyborohydride (126 mg, 0.294 mmol) was added. The mixture was stirred overnight. The mixture was filtered through glass fiber filter paper and washed 2× with 0.3 mL of DCE. Trifluoroacetic acid (0.18 mL, 2.336 mmol) was added and the mixture stirred at room temperature overnight. The reaction mixture was concentrated and taken up in 1.3 mL of DMSO and purified on a Gilson HPLC eluting at 20 mL/min with a linear gradient running from 10% CH₃CN/H₂O (0.1% TFA) to 90% CH₃CN/H₂O (0.1% TFA). The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking up the residue in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The organic phase was washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-{[5-chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (21.9 mg, 49.6%) as a white solid. LC-MS m/z 749 M⁺; mp 119-122° C.

Example 425 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic acid (37 mg, 0.079 mmol) was partially dissolved in the dichloromethane (DCM) (2 mL) and HBTU (30.1 mg, 0.079 mmol) was added followed sequentially by ({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)amine (23.40 mg, 0.079 mmol) and TEA (0.022 mL, 0.159 mmol). The mixture was allowed to stir under argon at room temperature overnight. Then more HBTU (3 mg, 0.0079 mmol) was added followed by ({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)amine (2.3 mg, 0.0079 mmol), and the reaction mixture stirred overnight again. The solvents were pumped off, and the residue taken up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The organic phase was washed with water (2×5 mL) and then with brine (1×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give the crude residue. It was taken up in DMSO (1 mL) and purified on a Gilson HPLC eluting at 20 mL/min with a linear gradient running from 0% CH₃CN/H₂O (0.1% TFA) to 90% CH₃CN/H₂O (0.1% TFA). The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated and the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide (30.6 mg, 48.6%) as an off-white solid. LC-MS m/z 743 (M+H)⁺; mp 119-121° C.

Example 426 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

3-[({[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)carbonyl]-5-methylbenzoic acid (37 mg, 0.079 mmol) was partially dissolved in the dichloromethane (DCM) (2.0 mL), HBTU (30.1 mg, 0.079 mmol) was added followed sequentially by 1,1-dimethylethyl 4-{[3′-(aminomethyl)-3-biphenylyl]methyl}-1-piperidinecarboxylate (30.2 mg, 0.079 mmol), and TEA (0.022 mL, 0.159 mmol). The mixture was allowed to stir under argon at room temperature overnight. The crude product was partitioned between EtOAc (30 mL) and water (10 mL). The phases were separated, and the organic phase washed with brine, dried over anhydrous Na₂SO₄ filtered and concentrated to give the crude residue. It was taken up in methylene chloride and absorbed on Isolute® Sorbent and purified on a 4 g silica column eluted with 0-10% MeOH/DCM. The product fractions were concentrated and taken up in DCM (1.8 mL), and then treated with trifluoroacetic acid (0.2 mL). The mixture was stirred under argon for 5 h. Solvents were pumped off and the residue taken up in DMSO and purified on the Gilson with the TFA system. The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated, the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide (31.91 mg, 0.043 mmol, 54.5% yield) as a white solid. LC-MS m/z 728 (M+H)⁺; mp 130-132° C.

Example 427 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({6-(methyloxy)-3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

N-{[3-Bromo-4-(methyloxy)phenyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide (50 mg, 0.075 mmol), 1-methyl-4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}piperidine (31.5 mg, 0.1 mmol), potassium carbonate (31.2 mg, 0.226 mmol), and Pd(Ph₃P)₄ (4.35 mg, 3.77 μmol) were added to a 0.5-2 mL Biotage microwave vial in 1,4-dioxane (1.5 mL) and water (0.5 mL). The vial was capped and the mixture was heated under microwave at normal power in the Emrys Optimizer at 100° C. for 15 min. The crude product was partitioned between EtOAc (30 mL) and water (10 mL). The phases were separated, and the organic washed with brine, dried over anhydrous Na₂SO₄ filtered and concentrated to give the crude residue. It was taken up in DMSO and purified on the Gilson with the TFA system. The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated, the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({6-(methyloxy)-3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide (34.1 mg, 58.2%) as a white solid. LC-MS m/z 772 (M+H)⁺; mp 115-117° C.

Example 428 N-({5-Chloro-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(5-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (34.0 mg, 0.05 mmol), 1-methylhexahydro-1H-1,4-diazepine (11.42 mg, 0.100 mmol), MP-Triacetoxyborohydride (107 mg, 0.250 mmol), and acetic acid (3.15 μL, 0.055 mmol) were combined in dimethyl sulfoxide (DMSO) (0.5 mL) in a 1 dram vial and stirred at room temperature overnight. The mixture was filtered through glass fibre filter paper and washed 2× with 0.3 mL of DMSO. The combined filtrates were purified on the Gilson with the TFA system. The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated, the organic phase washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-({5-chloro-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (24.2 mg, 62.3%) as a white solid. LC-MS m/z 777 M⁺; mp 111-113° C.

Example 429 N-[(5-Chloro-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(5-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (34.0 mg, 0.05 mmol), 2,6-dimethylpiperazine (11.42 mg, 0.100 mmol), and acetic acid (3.15 μL, 0.055 mmol) were combined in dimethyl sulfoxide (DMSO) (0.5 mL) in a 1 dram vial and stirred at room temperature for 30 min. The MP-Triacetoxyborohydride (107 mg, 0.250 mmol) was added and the mixture stirred overnight. The mixture was filtered through glass fiber filter paper and washed 2× with 0.3 mL of DMSO. The combined filtrates were purified on the Gilson with the TFA system. The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking the residue up in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The phases were separated, the organic phase was washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-[(5-chloro-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (25.6 mg, 65.9%) as a white solid. LC-MS m/z 777 M⁺; mp 112-114° C.

Example 430 N-{[5-Chloro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(5-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (40 mg, 0.059 mmol) and 1,1-dimethylethyl hexahydro-1H-1,4-diazepine-1-carboxylate (23.59 mg, 0.118 mmol) were dissolved in 1,2-dichloroethane (DCE) (1 mL) in a 1 dram vial and acetic acid (3.71 μL, 0.065 mmol) was added. The mixture was stirred for 30 min and then MP-Triacetoxyborohydride (126 mg, 0.294 mmol) was added. The mixture was stirred overnight. The mixture was filtered through glass fibre filter paper and washed 2× with 0.3 mL of DCE. Trifluoroacetic acid (0.18 mL, 2.336 mmol) was added and the mixture stirred at room temperature overnight. The reaction mixture was concentrated and taken up in 1.3 mL of DMSO and purified on a Gilson HPLC eluting at 20 mL/min with a linear gradient running from 10% CH₃CN/H₂O (0.1% TFA) to 90% CH₃CN/H₂O (0.1% TFA). The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking up the residue in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The organic phase was washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give N-{[5-chloro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (28.2 mg, 62.3%) as a white solid. LC-MS m/z 763 M⁺; mp 121-124° C.

Example 431 N-({5-Chloro-3′-[(1S,4S)-2,5-diazabicyclo [2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide

N-[(5-Chloro-3′-formyl-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (40 mg, 0.059 mmol) and 1,1-dimethylethyl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (11.68 mg, 0.059 mmol) were dissolved in 1,2-dichloroethane (DCE) (1 mL) in a 1 dram vial and acetic acid (3.71 μL, 0.065 mmol) was added. The mixture was stirred for 30 min and then MP-Triacetoxyborohydride (126 mg, 0.294 mmol) was added. The mixture was stirred overnight. The mixture was filtered through glass fibre filter paper and washed 2× with 0.3 mL of DCE. Trifluoroacetic acid (0.18 mL, 2.336 mmol) was added and the mixture stirred at room temperature overnight. The reaction mixture was concentrated and taken up in 1.1 mL of DMSO and purified on a Gilson HPLC eluting at 20 mL/min with a linear gradient running from 10% CH₃CN/H₂O (0.1% TFA) to 90% CH₃CN/H₂O (0.1% TFA). The desired fractions were concentrated to give the desired product as a TFA salt. This was converted to the free base by taking up the residue in EtOAc (30 mL) and 2.5 N NaOH (5 mL). The organic phase was washed with 2.5 N NaOH (3×5 mL) and then with brine (2×5 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and dried under 50° C. to give N-({5-chloro-3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (25.8 mg, 57.5%) as a white solid. LC-MS m/z 762 (M+H)⁺; mp 130-133° C.

Example 432 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide

To a solution of N-[(3-bromo-4-fluorophenyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide (0.641 g, 0.1 mmol) in 1,4-dioxane (1.5 ml) and water (0.500 ml) was added 1,1-dimethylethyl 4-{[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}-1-piperidinecarboxylate (0.048 g, 0.120 mmol), Pd(Ph₃P)₄ (4.62 mg, 4.00 μmol), and K₂CO₃ (0.041 g, 0.300 mmol). This mixture was heated using a microwave instrument at 150° C. for 15 min. The organic layer was separated, concentrated using a Glas-Col evaporator, purified using a Gilson HPLC (with TFA), concentrated by GeneVav EZ-2 evaporator. The resulting solid was added to DCM (0.3 mL) then TFA (0.15 mL) and this mixture was left stand for 30 min before it was concentrated using a Glas-Col evaporator. The concentrate was purified using a Gilson HPLC (with TFA), concentrated by GeneVav EZ-2 evaporator, basified by amine cartridge to afford the title compound 0.016 g (21%). LC-MS m/z 732 (M+H)⁺.

Example 433 N-{[1,6-Diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(3,4-dimethyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-1,3-benzenedicarboxamide

A mixture of N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-formyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide (150 mg, 0.226 mmol), 1,2-dimethylpiperazine (45.7 mg, 0.40 mmol), sodium triacetoxyborohydride (95 mg, 0.45 mmol) and AcOH (18 μl, 0.314 mmol) in 1,2-dichloroethane (DCE) (6 mL) was stirred overnight at room temperature. LC/MS showed that the reaction was complete. The reaction was diluted with EtOAc; the organic layer washed with dilute aq. NaOH, then sat. aq. NaCl and dried [Na₂SO₄], then concentrated to a viscous taffy. The material was purified on a Gilson HPLC using reverse phase (acetonitrile-water with 0.1% TFA). Fractions containing the desired product were combined, the MeOH removed, the aqueous layer treated with EtOAc and 1N NaOH. The mixture was vigorously extracted with EtOAc (2×), the extract was washed with a small amount of sat. aq. NaCl; dried [Na₂SO₄], then concentrated to a glassy, creamy tan, coloured foam (0.1002 g). LC-MS m/z 761.5 (M+H)⁺, 0.76 min (ret time).

All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth.

The above description fully discloses the invention including preferred embodiments thereof. Modifications and improvements of the embodiments specifically disclosed herein are within the scope of the following claims. Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. Therefore, the Examples herein are to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows. 

1. A compound which is selected from the group consisting of: N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro -3′-(4-morpholinylmethyl)-3-biphenylyl]methyl -1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl }-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-6-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro -1H-1,4-diazepin-1-ylmethyl)-6-methyl-3-biphenylyl]methyl 1-1,3-benzenedicarboxamide; N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-6-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(2R,5S)-2,5-dimethyl-1-piperazinyl]methyl }-6-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro -3′-(1-piperidinylmethyl)-3-biphenylyl]methyl }-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(3-oxo-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(4-ethyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(1-methylethyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[[2-(dimethylamino)-ethyl](methyl)amino]methyl}-6-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S )-3,5-dimethyl-1-piperazinyl]methyl}-6-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-methyl-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-[(6-chloro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-methyl-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-({6-chloro-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-1,3-benzenedicarboxamide; N-({6-chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl 1methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-[(6-chloro-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[6-chloro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[6-chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-({6-chloro-3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-[(5-chloro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; 3-{[{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}(methyl)amino]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}benzamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-{[{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}(methyl)amino]methyl}benzamide; 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)methyl]-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}benzamide; 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}oxy) methyl]-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}benzamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-fluoro-3′-[(4-methyl-1-piperazinyl) methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N,N′-dimethyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N,N′-dimethyl -1,3-benzenedicarbox amide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,4-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(2-oxo-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-(methyloxy)-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-5-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin -5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-methyl-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-5-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S )-3,5-dimethyl-1-piperazinyl]methyl}-5-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-5-methyl-3-biphenylyl}methyl)-N -{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(4-methyl-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-4-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S )-3,5-dimethyl-1-piperazinyl]methyl}-4-methyl-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[4-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({4-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-4-methyl-3-biphenylyl}methyl)-N -{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(5-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3-fluorophenyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-5-fluoro-3-biphenylyl}methyl)-N -{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[5-fluoro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-5-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({5-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[6-cyano-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N -{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-[(6-cyano-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-({6-cyano-3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[6-cyano-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-[(6-cyano-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-({6-cyano-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-({6-cyano-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[4-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-4-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin -5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({4-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-4-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(4-(methyloxy)-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[2-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-({3′-[{(1S,4S)-2,5-ciazabicyclo[2.2.1]hept-2-ylmethyl]-2-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-2-methyl-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S )-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-methyl-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(2-methyl-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3R)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(2-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′4(3,5-dimethyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(2-hydroxyethyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(2,3,5,6-tetramethyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({4-[2-(methyloxy)ethyl]-1-piperazinyllmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({442-(dimethylamino)ethyl]-1-piperazinyl}methyl)-6-fluoro-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-phenyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(4-pyridinyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({methyl [2-(methylamino)ethyl]amino 1methyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-[(3′-{[(cyclopropylmethyl)amino]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl -4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[methyl(1-methyl-4-piperidinyl)amino]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(2,2,6,6-tetramethyl-4-piperidinyl)amino]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(4-morpholinyl)-1-piperidinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-({3′-[(1-azabicyclo[2.2.2]oct-3-ylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-V′-[(3′-{[(1-ethyl-3-piperidinyl)amino]methyl}-6-fluoro-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({[(3R)-2-oxohexahydro-1H-azepin-3-yl]aminolmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-[(3′-{[(25)-2-(amino carbonyl)-1-pyn⁻olidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N -{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({2-[(dimethylamino)methyl]-1-piperidinyl}methyl)-6-fluoro-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-[(3′-{[3-(aminocarbonyl)-1-piperidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(2-pyrimidinyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-({3′-[(4-acetyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(1R,4S)-5-(phenylmethyl)-2,5-diazabicyclo[2.2.1]hept-2-yl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-({4-[(3-methylphenyl)methyl]-1-piperazinyllmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[4-(4-fluorophenyl)-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-({3′-[(cyclopentylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-1,3-benzenedicarboxamide; N-({3′-[(cyclohexylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-N′-({6-fluoro -3′-[(2-pyridinylamino)methyl]-3-biphenylyl 1methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-N′-({6-fluoro -3′-[(3-pyridinylamino)methyl]-3-biphenylyl 1methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro -3′-[(4-pyridinylamino)methyl]-3-biphenylyl 1methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro -3′-[(4-pyrimidinylamino) methyl]-3-biphenylyl 1methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro -3′-[(2-pyrimidinylamino) methyl]-3-biphenylyl 1methyl)-1,3-benzenedicarboxamide; N-({3′-[(diethylamino)methyl]-6-fluoro-3-biphenylyl}methyl)-′-{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-1) ]pyridin-5-yl]methyl 1-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({[2-(dimethylamino)ethyl]aminol-methyl)-6-fluoro-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-[(3′-{[3-(aminocarbonyl)-2,2,5,5-tetramethyl-1-pyrrolidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin -5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl 1methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-piperidinylamino)methyl]-3-biphenylyl 1methyl)-1,3-benzenedicarboxamide; N-({3′-[(4-amino-1-piperidinyl)methyl]-6-fluoro-3-biphenylyl 1methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-[(3′-{[(3S )-3-amino-1-pyn⁻olidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(4-piperidinylmethyl)amino]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(hexahydro-1H-azepin-3-ylamino)methyl]-3-biphenylyl }methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(hexahydro-1H-azepin-4-ylamino)methyl]-3-biphenylyl }methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl }-3-biphenylyl)methyl]-2,3-benzenedicarboxamide;naphthalenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl }-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-hydroxy-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-(1 , 1-dimethylethyl)-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-2,4,5,6-tetrafluoro-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl }-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; 5-bromo-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl }-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-naphthalenedicarboxamide ; 2,5-dichloro-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin -5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-2,3,5,6-tetrafluoro-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,4-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-({ethyl [2-(methylamino)ethyl]amino Imethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,4-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-hydroxy-1-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′-[(4-oxo-1-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′44-piperidinyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-({3′-[(4-cyano-1-piperidinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-N′-methyl-1,3-benzenedicarboxamide; 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]benzamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-[({[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}amino)sulfonyl]benzamide; 2-chloro-5-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-4-fluoro-N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]b enzamide; 5-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-2-(methyloxy)benzamide; 3-{4-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)sulfonyl]phenyll -N′-[(6-fluoro-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]propanamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-[(methyl {[3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}amino)sulfonyl]benzamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-({methyl [(3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]amino 1sulfonyl)benzamide; 3-[({[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}amino)methyl]-N′-[(4′-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]benzamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-3-{[({3′4(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)amino]sulfonyllbenzamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(8-methyl-8-azabicyclo[3.2.1 ]oct-3-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[2-(3-{[(3S)-3-methyl-1-piperazinyl]methyl}phenyl)-4-pyridinyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({2-[3-(4-piperidinylmethyl)phenyl]-4-pyridinyllmethyl)-1,3-benzenedicarboxamide; N-[(3′-{(1S,4S)-5-Acetyl-2,5-diazabicyclo[2.2.2]oct-2-yl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin -5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(3R)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-({3′4(1S,4S)-2,5-diazabicyclo[2 .2.1 ]hept-2-ylmethyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl -1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-[(3′-{[(3S)-3-amino-1-pyrrolidinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-N′-{[1,6-benzenedicarboxamide;diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(6-fluoro-3′-{[(2S)-2-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[3′41-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(3′-{[(3R)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-({3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(3′-{[(2S)-2-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[6-chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-[(6-chloro-3′-{[(3R)-3-methyl-1-piperazinyl]methyl 1-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-({6-chloro-3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-3-biphenylyl 1methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl -1,3-benzenedicarboxamide; N-[(6-chloro-3′-{[(3S )-3-methyl-1-piperazinyl]methyl 1-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[6-chloro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)- 3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-[(3′-{[(3S )-3-amino-1-pyrrolidinyl]methyl}-6-chloro-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-[(6-chloro-3′-{[(25)-2-methyl-1-piperazinyl]methyl 1-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-({6-chloro-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl 1methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-5-methyl-1,3-benzenedicarboxamide; N-({6-chloro-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl 1methyl) -N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-5-methyl-1,3-benzenedicarboxamide; N-({3′-[(4-acetyl-1-piperazinyl)methyl]-6-chloro-3-biphenylyl 1methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-5-methyl-1,3-benzenedicarboxamide; N-({6-chloro-3′-[(4-ethyl-1-piperazinyl)methyl]-3-biphenylyl 1methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-5-methyl-1,3-benzenedicarboxamide; N-[(6-chloro-3′-{[[2-(dimethylamino)ethyl](methyl)amino]methyl 1-3-biphenylyl)methyl]-N -{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl -1,3-benzenedicarboxamide; N-{[6-chloro-3′-(1-piperidinylmethyl)-3-biphenylyl]methyl 1-N′-{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl 1-5-methyl-1,3-benzenedicarboxamide; N-[(6-chloro-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′4(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({6-fluoro-3′4(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-5-methyl -1,3-benzenedicarboxamide; N-({3′-[(4-acetyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(4-ethyl-1-piperazinyl)methyl]-6-fluoro-3-biphenylyl}methyl)-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[[2-(dimethylamino)ethyl](methyl)amino]methyl}-6-fluoro-3-biphenylyl)methyl]-5-methyl -1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(1-piperidinylmethyl)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-6-fluoro-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-({3′-[(4-acetyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(4-ethyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′4(3′-{[[2-(dimethylamino)ethyl](methyl)amino]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[3′-(1-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-[(3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-(methyloxy)-3′-{[(3R)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-methyl-3′-{[(3R)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-methyl-3′-{[(3S )-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[3′-[(1R,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-6-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin -5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-(methyloxy)-3′-{[(2S)-2-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-6-methyl-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-[(3′-{[(3S )-3-amino-1-pyrrolidinyl]methyl}-6-methyl-3-biphenylyl)methyl]-N′-{{1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-1)]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[3′-(2,5-diazabicyclo[2.2.1]hept-2-ylmethyl)-6-methyl-3-biphenylyl]methyl 1-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-methyl-3′-{[(25)-2-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[6-(methyloxy)-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[6-methyl-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({6-(methyloxy)-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({6-methyl-3′-[(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-6-(methyloxy)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[3′-[(4-acetyl-1-piperazinyl)methyl]-6-(methyloxy)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-[(4-ethyl-1-piperazinyl)methyl]-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[[2-(dimethylamino)ethyl](methyl)amino]methyl}-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[6-(methyloxy)-3′-(1-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-6-(methyloxy)-3-biphenylyl]methyl}-5-methyl -1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({6-methyl-3′-[(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl) -1,3-benzenedicarboxamide; N-({3′-[(4-acetyl-1-piperazinyl)methyl]-6-methyl-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-({3′-[(4-ethyl-1-piperazinyl)methyl]-6-methyl-3-biphenylyl}methyl)-5-methyl-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′4(3′-{[[2-(dimethylamino)ethyl](methyl)amino]methyl}-6-methyl-3-biphenylyl)methyl]-5-methyl -1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[6-methyl-3′-(1-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′4(3′-{[(3R,5S )-3,5-dimethyl-1-piperazinyl]methyl}-6-methyl-3-biphenylyl)methyl]-5-methyl-1,3-benzenedicarboxamide; N-({5-chloro-3′4(4-methyl-1-piperazinyl)methyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-[(6-(methyloxy)-3′-{[(3S)-3-methyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-1,3-benzenedicarboxamide; N-{[5-chloro-3′-(1-piperazinylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro -2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({3′4(1-methyl-4-piperidinyl)methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-{[3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-5-methyl-N′-({6-(methyloxy)-3′4(1-methyl-4-piperidinyl) methyl]-3-biphenylyl}methyl)-1,3-benzenedicarboxamide; N-({5-chloro-3′4(4-methylhexahydro-1H-1,4-diazepin-1-yl)methyl]-3-biphenylyl}methyl) -N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-[(5-chloro-3′-{[(3R,5S)-3,5-dimethyl-1-piperazinyl]methyl}-3-biphenylyl)methyl]-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[5-chloro-3′-(hexahydro-1H-1,4-diazepin-1-ylmethyl)-3-biphenylyl]methyl}-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-({5-chloro-3′-[(1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-ylmethyl]-3-biphenylyl}methyl)-N′-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-1,3-benzenedicarboxamide; N-{[1,6-diethyl-4-(tetrahydro-2H-pyran-4-ylamino)-1H-pyrazolo[3,4-b]pyridin-5-yl]methyl}-N′-{[6-fluoro-3′-(4-piperidinylmethyl)-3-biphenylyl]methyl}-1,3-benzenedicarboxamide; and a pharmaceutically acceptable salt thereof.
 2. A pharmaceutical composition comprising a compound a compound according to claim 1, and a pharmaceutically acceptable carrier or diluent.
 3. A method of treating asthma, chronic obstructive pulmonary disease or rhinitis in a patient in need thereof, comprising administering to said patient an effective amount of a compound according to claim
 1. 